Cell Biology Product and Research Guide PDF

2013 Cell biology
product and resource guide
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Cell biology resources
2013 Life Technologies
Cell biology product and resource guide

Cell biology products
Comprising trusted and familiar Gibco, Invitrogen, Molecular Probes, and Novex products, the Life Technologies cell biology
portfolio offers published, high-performance, scalable solutions across the breadth of cell biology applications.
The book begins with overviews of five prominent cell biology research areas: neurobiology, immunology, cancer, stem cell biology,
and plant cell biology. Using published data, weve also constructed pathway maps for some of these research areas that you may
consult as you plan your research.
Sample
preparation
Key
technologies
Essentials
Cell culture and cells (Chapter 2)
Cell analysis (Chapter 3)
Recovery Freezing Medium, page 40

Gibco RPMI Media 1640, page 41
Gibco Minimal Essential Medium (MEM), page 41
GlutaMAX media, page 41
Opti-MEM reduced-serum medium, page 41
Dynabeads cell isolation technology, page 76

Dynabeads CD3/CD28 technology, page 79
Expi293 Expression System, page 42

FreeStyle MAX CHO Expression system, page 42
AlgiMatrix 3D cell culture system, page 49
Geltrex LDEV-free matrix products, page 49
AmnioMAX media, page 51
MarrowMAX bone marrow medium, page 51
PB-MAX Karyotyping Medium, page 51
KaryoMAX medium, page 51
TrypLE reagent, page 52
Click-iT EdU cell proliferation assay, page 82

Premo FUCCI Cell Cycle Sensor, page 83
CellROX Deep Red Reagent, page 86
CellLight fluorescent protein constructs, page 87
Qtracker Cell Labeling, page 95
FluoroMyelin Green fluorescent myelin stain, page 95
Molecular Probes antibodies for multicolor flow
Classical Gibco media: DMEM, RPMI, and MEM,
DynaMag magnets, page 80

HulaMixer Sample Mixer, page 80
LIVE/DEAD Viability/Cytotoxicity Kits, page 81
CellEvent Caspase-3/7 Green Detection Reagent,
page 41
Advanced Gibco media: GlutaMAX, OptiMEM,
page 41
Gibco FBS, page 43
Geneticin selection antibiotic, page 46
Puromycin seletion antibiotic, page 46
Zeocin selection antibiotic, page 46
Gibco growth factors, page 47
B-27 Supplement, serum substitute for neural cell
culture, page 72
Neurobasal Media, basal media, page 48
KnockOut Serum Replacement, for pluripotent stem
cells, page 60
cytometry, page 102
Qdot nanocrystals for flow cytometry, page 105

Zenon antibody labeling, page 107
APEX antibody labeling, page 107
page 85
pHrodo indicators, page 87

Alexa Fluor phalloidins, page 90
MitoTracker dyes, page 91
JC-1 dye, page 91
LysoTracker dyes, page 92
BODIPY FL C5-ceramide, page 93
TO-PRO-3, page 94
Calcein, AM, page 95
CellTracker dyes, page 95
Fluorescent dextrans, page 95
Alexa Fluor conjugated secondary antibodies and
streptavidin, page 106
Detection
instruments
Countess Automated Cell Counter, page 53

Neon Transfection System, page 55
Tali Image-Based Cytometer, page 55
FLoid Cell Imaging Station, page 55
Attune Acoustic Focusing Cytometer, page 55
Tali Image-Based Cytometer, page 99

FLoid Cell Imaging Station, page 101
Attune Acoustic Focusing Cytometer, page 100
2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.
Chapter 1 contains tools you may find useful to choose products and interpret your data.
Cell biology products
The rest of the guide is summarized in the chart belowan organized snapshot of the most popular of our reliable solutions,
backed by nearly 3 million publications. For complete product information, refer to the chapter/page numbers noted.
Protein analysis (Chapter 4)

Dynabeads immunoprecipitation, page 129
Novex organelle isolation kits, page 132
Novex protein A/G purification, page 130
Novex cell lysis reagents, page 133
Protein expression
Champion pET Expression System, page 119

PichiaPink Yeast Expression System, page 118
GeneArt Algae Engineering Kit, page 120
GeneArt Gene Synthesis, page 121
Lipofectamine LTX Transfection Reagent, page 123
Lipofectamine RNAiMAX Reagent, page 123
Lipofectamine 2000 Transfection Reagent, page 123
Silencer Select siRNA, page 126
Stealth Select siRNA and Silencer siRNA, page 126
mirVana miRNA Mimics and Inhibitors, page 128
Protein expression
Ambion Pre-miR Precursors and Anti-miR Inhibitors,

page 128
Lipofectamine reagents, page 123
Qubit Fluorometer, page 134

Neon Transfection System, page 124
POROS analytical chromatography, page 131
ZOOM IEF Fractionator, page 132
Protein analysis
Bolt and NuPAGE protein gels, page 136

Novex Sharp and SeeBlue Plus 2 Pre-Stained Standard,
page 141
SYPRO Ruby Protein Gel Stain, page 142

Novex ABfinity antibodies, page 147
Novex multiplex immunoassays for Luminex platforms, page 153
Novex magnetic multiplex immunoassay for Luminex, page 154
Protein analysis
Novex pour-your-own gel essentials, page 136

Novex Tris-glycine gels, page 136
SeeBlue Pre-Stained Standard, page 141
SimplyBlue SafeStain, page 142
Novex Antibody Pair kits, page 150
Novex ELISA kits, page 150
XCell SureLock Protein Electrophoresis Cells, page 139

iBlot 7-Minute Blotting System, page 143
BenchPro 4100 Western Processing System, page 144
MAGPIX System, page 155
lifetechnologies.com
0
1
2
3
4
Cell biology research and pathways
Cell culture and cells
Cell analysis
Protein analysis
The products listed in this document have the following product uses.
Product type
Product Use
Research use
For Research Use Only. Not for use in diagnostic procedures.
Class I - IVD
In vitro Diagnostic Use
CTS brand research use
For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. CAUTION: Not intended for direct administration into human or animals.
OR
For Research Use or Non-Commercial Manufacturing of Cell-Based Products for Clinical Research. Caution: Not intended for direct administration
into humans or animals.
Class II medical device
For human ex vivo tissue and cell culture processing applications: CAUTION: When used as a medical device, Federal Law restricts this device to sale
by or on the order of a physician.
Unless otherwise indicated, the products included in this catalog are For Research Use Only. Not for use in diagnostic procedures.

Research areasThis section provides an overview for investigators in a
specific discipline to see the broad menu of products, services, and tools
Life Technologies offers for their research and tools.
Neurobiology research
Immunology research
14
Cancer research
16
Stem cell research
18
Plant cell biology research
20
Cellular signaling pathwaysRefer to these publication-supported

representations of several important signaling pathways.
AKT pathway
22
MAPK pathway
24
JAK/STAT pathway
26
Neurodegeneration 28
Custom servicesTake advantage of these terrific service
offerings to customize your tools.
Gibco custom media services
30
Custom Novex multiplex assays for Luminex technology
32
Custom biology services
33
Website resourcesAll of these resources are available at a click, including

resources for learning techniques, finding the right cell line by application,
downloading protocols, stain and antibody selection guides, and mobile apps
for the scientist on the go.
Gibco Cell Culture Basics
34
Cell Line Database
34
Protocol Database
35
The Molecular Probes Handbook online
35
Fluorescence SpectraViewer
36
Mobile applications and widgets
36
Virtual Cell Staining Tool
36
Antibody and immunoassay selection guides
37
Customer and technical support
37
Life Technologies offers a range of cells and cell culture tools as well as cellular analysis assays and
reagents for research on neural primary and stem cells. Our products encompass all major neural cell
types and support a multitude of applications from basic research to therapeutic discovery. Select the right
tools for your neuroscience research at lifetechnologies.com/neuro.
Cells and culture tools for neurobiology

Neural primary and stem cells
Gibco offers a wide array of highly pure, ready-to-use primary and stem cells for neural research that
have high viability and superior performance. For more details on cells available, visit lifetechnologies.com/
neuroculture.
Neural primary cells:
Rat cortex and hippocampus neuronal cells

Rat primary cortical astrocytes
Human astrocytes
Stem cells:
Human neural stem cells
Rat fetal neural stem cells
Rat glial precursor cells
Human dopaminergic precursor cells (available as custom order only)
Primary rat hippocampus neurons
Human neural stem

cells
Media and supplements

Selecting the right media system for your neural cell culture is important to ensure experimental consistency, reproducibility, and relevancy. For over twenty years, neuroscience researchers have relied on Gibco
products to ensure confidence in results. Cited in over 11,000 publications spanning more than 50 cell types
and an expansive list of disease models, Gibco neural cell culture products deliver the versatility required
from media systems today.
Choose from supplements optimized to support a variety of applications, basal media designed for key cell
types, and quality primary and stem cells. To learn more, visit lifetechnologies.com/neuroculture.
To build the optimal media system for your neural cell culture needs, explore our product offering by cell
type, or browse our products designed for specific customer applications like electrophysiology or insulin
studies, in the tables on pages 9 and 10.

Tools for primary neurons.
Basal media
Optimized cell type
Cells
Cell source
Growth and
maintenance of
neurons
Neurobasal
Medium
Optimized for
prenatal and fetal
neurons
Primary rat cortex

neurons
Isolated from day-18

Fisher 344 rat
embryos
Neurobasal-A
Medium
Optimized for
postnatal and adult
brain neurons
Primary rat
hippocampus
neurons
Isolated from day-18

Fisher 344 rat
embryos
DMEM media
Growth of a
spectrum of
mammalian cell
lines
Astrocyte Medium
Growth of primary
and serial tumor
lines of astrocytic
phenotype
Rat primary cortical

astrocytes
Isolated from
cortices of SpragueDawley rats at
embryonic day 18
(E19)
Growth of primary
human and rat
astrocytes
Human astrocytes
Normal human cells

derived from human
brain tissue
Primary neurons
Application
B-27 Serum Free

Supplement
Glial
cells
G-5 Supplement
Supplement
Growth of glial cell

lines
Astrocytes
G-5 Supplement
Progenitor cells
Neural stem cells
Tools for neural stem cells and progenitor cells.

Supplement
Application
Basal media
Optimized cell type
Cells
Cell source
B-27 Supplement,
minus Vitamin A
Proliferation of stem
cells
StemPro NSC SFM
Human neural stem

cells
Human neural stem

cells
Derived from H9
(WA09) human
embryonic stem
cells
B-27 Supplement,
XenoFree
Differentiation of
stem cells
KnockOut DMEM/
F-12 Medium
Neural stem cells
Rat fetal neural

stem cells
Isolated from the

cortex of SpragueDawley E14 rats
N-2 Supplement
Serum substitute for

embryonic neural
cells
StemPro Neural
Supplement
Growth and expansion of human and

rat neural stem
cells (NSCs)
N-2 Supplement
Serum substitute
for CNS progenitor
cells
KnockOut DMEM/
F-12 Medium
Neural stem cells
Rat glial precursor

cells (rGPCs)
Isolated from
newborn SpragueDawley rat cortex
StemPro Neural
Supplement
Growth and expansion of progenitor

cells
Human dopaminergic precursor

cells*
Derived from H9
human embryonic
stem cells (ESCs)
* Available as custom order.
Tools optimized for specialty applications.

Supplement
Application
B-27 Supplement, minus AO (antioxidants)
Studies of oxidative stress/damage,

apoptosis, or where free radical damage to
neurons occurs
B-27 Supplement, minus insulin
Studies of insulin secretion or insulin

receptors
B-27 Electrophysiology Kit
Electrophysiology studies
B-27 Supplement, custom
Growth and expansion of retinal ganglion

cells
Basal media
Application
Optimized cell type
Neurobasal Medium without phenol red
Receptor studies, downstream purification

studies, or other processes where the presence of phenol red is undesired
Optimized for postnatal and adult brain

neurons
Neurobasal-A Medium without phenol red

Hibernate-A Medium
Manipulation of neurons at ambient CO2 for at

least 48 hr while retaining viability
Hibernate -E Medium
Prenatal and fetal neurons
Optimized for postnatal neurons

Optimized for embryonic neurons
Serum-free supplements
B-27 Supplementsoptimized serum substitute for neural cell culture available exclusively from Life
Technologies; learn more at lifetechnologies.com/b27
N-2 Supplementserum substitute for embryonic neural and central nervous system progenitor cells
G-5 Supplementchemically defined, serum-free supplement for growth of glial cell lines
StemPro Neural Supplementserum-free supplement formulated for the growth and expansion of human
and rat NSCs and progenitor cells
Gibco seradeliver consistent cell growth over time and passages; learn more at lifetechnologies.com/
cellculture
Supplement comparison.
10
Standard for mammalian cells
Industry standard for neuronal cell

culture
Optimized for neuronal stem cells
Product
Gibco sera
B-27 Serum Free Supplement
StemPro Neural Supplement &

B-27 Supplement, XenoFree
Lot-to-lot consistency
Highest grade only
Yes
Yes
cGMP manufacturing
Yes
Yes
Yes
Performance testing
Non-neural cell line proliferation
Primary neuron cell survival
Proliferation and differentiation of

human NSCs
Compatible cell types
Neural and non-neural cell lines
Human and animal primary neurons

and glia
Human and animal NSCs
Formulations offered
Qualified and certified
Minus antioxidants, minus insulin, or

electrophysiology
Minus vitamin A and XenoFree
Learn more
lifetechnologies.com/cellculture
lifetechnologies.com/b27
lifetechnologies.com/NSC

Basal media
Neurobasal mediasupport growth of neurons when combined with B-27,
N-2, or G-5 supplements and designed to inhibit glial proliferation; learn more at
lifetechnologies.com/neurobasal
KnockOut DMEM/F-12 Mediumoptimized for growth of undifferentiated stem
cells; supports growth of NSCs when used with StemPro Neural Supplement
StemPro NSC SFMserum-free medium formulated for growth and expansion
of human NSCs derived from either ESCs or fetal tissue; kit includes KnockOut
DMEM/F-12 medium, StemPro Neural Supplement, and growth factors
Gibco DMEMClassic formulation used to culture glial cells (astrocytes and
oligodendrocytes)
Cell substrates
Natural mouse lamininimportant for cell attachment, spreading, and neurite

outgrowth
Fibronectinmodulates cell adhesion, growth, migration, and differentiation as a

component of the extracellular matrix
CELLstart CTS Substratefully defined, xeno-free substrate for stem cell

growth and expansion; maintains multipotency of human NSCs in StemPro
NSC SFM
Enzymatic passaging
StemPro Accutase Cell Dissociation Reagentdesigned for gentle and efficient

detachment of neural stem cells, progenitor cells (including glial), and primary
neural cells
Cryopreservation
Synth-a-Freeze Cryopreservation Mediuma defined liquid cryopreservation

medium for human keratinocytes, ESCs, NSCs, and mesenchymal stem cells
Hibernate mediaideal for manipulation of neurons at ambient CO2 and as preservation media for viable brain tissue for up to a month at 4C
11

Cellular analysis assays and reagents for neurobiology
Cell viability assays
We have a range of cell viability assays to assess the health of your neural cells.
View more information and compare cell viability assays at lifetechnologies.com/cellhealthneuro.
Types of cell viability assays.

Fast resazurin-based
assays
Quantitative measurement
of DNA content
Quantitative measurement
of neurite outgrowth
Simultaneous quantification of live and dead cells
Product
PrestoBlue Cell Viability

Reagent
CyQUANT Cell Proliferation Assay
Neurite Outgrowth Staining

Kit
LIVE/DEAD cell viability

assays
Assay type
Single parameter
Single parameter
Multiplexed
Multiplexed
Mechanism of action
Metabolic activity
DNA quantification
Esterase activity,
membrane integrity
Esterase activity,
membrane integrity
Format
Live
Live
Live or endpoint
Endpoint
Incubation time
>10 min
3060 min
30 min
1545 min
Readout
Absorbance or fluorescence
Fluorescence
Fluorescence
Fluorescence
Instrument compatibility
Microplate reader
Microplate reader, microscopy
Microplate reader, microscopy
Flow cytometry, microplate

reader, microscopy
For more info, visit:
lifetechnologies.com/
prestoblue
cyquantdirect
neuriteoutgrowth
livedead
Fluorescent probes for neuronal physiology

Choose from a diverse selection of fluorescent ion indicators and probes for calcium, sodium, zinc, potassium,
magnesium, pH, membrane potential, and synaptic function from the industry leader in fluorescent technology.
FluxOR Potassium Ion Channel Assayan optically based, homogeneous assay for high-throughput
screening measurements of potassium ion channel and transporter activities; learn more at
lifetechnologies.com/fluxor
Fluo-4 Direct Calcium Assay Kitadvanced formulation that suppresses background fluorescence
generated from media with negligible impact on the cellular fluorescence, allowing the assay to be
run in a simple addition-only format in the presence of serum-containing media; learn more at
lifetechnologies.com/fluo4direct
Voltage sensor probesthese probes use a fluorescence resonance energy transfer-based voltage-sensing
assay technology to measure changes in cell membrane potential; learn more at lifetechnologies.com/vsp
FM probes for synaptic functionthese probes are ideal for investigating the mechanisms of synaptic
vesicle activity; learn more at lifetechnologies.com/fmprobes
Probes for neuronal tracing and anatomy
Choose from an extensive range of fluorescent and biotinylated tracers for retrograde and anterograde
tracing, fluorescent histological stains, and ligands for receptors, ion channels, and ion carriers.
Visit lifetechnologies.com/neuroprobes for a full list of probes for neuronal tracing and anatomy.
12

Detection of neuro-related proteins
Tools for neuro-related protein detection.
Novex product
Industry standard
Multiplexable
Western blotting
ELISA
Multiplex assay using Luminex

platform
Analytes per well
One
One
Up to 50 (proteins)
Hands-on time
23 days
35 hours
45 hours
Dynamic range
Low
Medium
High
For more info visit
Western products:
lifetechnologies.com/westerndetection
Antibodies:
lifetechnologies.com/abfinity
lifetechnologies.com/antibodies
lifetechnologies.com/neuroelisa
lifetechnologies.com/luminex
Qualitative
Cellular expression analysis

BacMam technology
BacMam technology allows simple, safe, efficient delivery and expression of genes of interest in mammalian
cells, including neurons.
For more information about BacMam technology, visit lifetechnologies.com/bacmam.
Cellular and pathway analysis
Target-based assays
Select from our broad range of validated cell-based assays for key neural targets, including G-protein
BacMam
reagentkinases,
construction
coupled
receptors,
and ion channels.
Visit lifetechnologies.com/discoveryassays.
Custom assay development
Let us create and validate a custom assay that meets your defined requirements.
Visit lifetechnologies.com/custom.
Target gene tranfer plasmid
Clone into baculovirus DNA
Transfect S9 insect cells
Transduce mammalian cells
Configure assays
Collect BacMam virus
Assay development and screeening
Use BacMam stocks
Perform screen or assay
BacMam workflow. Just add the BacMam reagent to your cells for 24 hours, treat with an enhancer (recommended for certain cell types including neurons), wash,
incubate overnight, and perform the assay.
13
Immunology research
Immunology research
At Life Technologies, we understand the importance of using the highest-quality and most sensitive reagents
to dissect the intricacies associated with immunology research. Our extensive portfolio of products can be used
across all stages of T and B cell development and activation, from isolation to stimulation and from expansion
to analysis.
Isolate: Dynal magnetic cell separation technologies yield pure,

uncontaminated cells without columns.
Stimulate: Our validated reagents allow better control using state-of-the-art

expansion technology and Gibco cytokines with consistently high bioactivity.
Ambion siRNA technology is ideal to knock down key genes and proteins
involved in immunology processes.
Analyze: We offer products for flow cytometry, imaging microscopy, western
blot analysis, and ELISA as well as Novex multiplex assays for protein
quantification using Luminex xMAP technology.
Check out our immunology-related signaling pathways on pages 15 and 22-29.
These include:
The inflammatory response

The AKT pathway
The MAPK pathway
The JAK/STAT pathway

Neurodegeneration
Find a comprehensive selection of our immunology products at lifetechnologies.com/immunology.
Below is a quick guide to the sections where you can learn more about immunology products and solutions from Life Technologies.
Isolate
Reagents
Dynabeads magnetic cell separation

products for:
Human cells
Mouse cells
Other cell types
Isolation of your own antibody cells
CD4+ cell counting kits
Primary antibodies for:
Human immunology
Mouse immunology
Rat immunology
Assays and kits
Positive isolation
Negative isolation
Depletion
Technologies
14
Instrumentation
Countess Automated Cell Counter
Accessories
Magnets and mixers
Stimulate
Specialty media for immunology research:
Gibco OpTmizer Cell Expansion
Serum-Free Medium
Gibco Serum-Free AIM V Medium
Gibco Macrophage Serum-Free Medium
Serum-free medium for hematopoietic
stem cell culture
Cytogenetic media
Recombinant growth factors and cytokines
Analyze
Primary antibodies for:
Human immunology
Mouse immunology
Rat immunology
Phosphorylation sitespecific primary
antibodies
Secondary detection reagents
Antibody pairs
Novex multiplex assays for the Luminex

platform
ELISA kits
Cell viability kits
Cell proliferation kits
Dynabeads technology for mouse and
human T cell activation and expansion
siRNA
Click chemistry
Luminex xMAP technology
CellLight reagents (BacMam technology)
ABfinity recombinant monoclonal antibody
technology
Neon Transfection System
Attune Acoustic Focusing Cytometer

FLoid Cell Imaging Station
Tali Image-Based Cytometer
MAGPIX System
iBlot Western Transfer System
Flow cytometry controls and standards
Imaging standards and calibration kits
Protein labeling kits
Chemiluminescence detection systems
Inflammation
PECAM-1, VCAM-1,
ICAM-1, P-selectin,
E-selectin
Bacterial products
(LPS, peptidoglycans, etc.)
Fibroblast
liver
Cardiovascular
pathology
M-CSF, TNF-, GM-CSF,

G-CSF, IL-1, IL-6, IL-11,
IL-12
am
st
Macrophage
Hematopoiesis
in
IL-8,
TNF-
Endothelial
cell
Hi
Fever
TNF, IL-1
Immunology research
TNF-
IL-1
TGF-
PDGF
IL-6
Acute phase
proteins
IL-1, IL-6, IL-8, IL-10,

IL-12, IL-15, IFN-, IFN-,
TNF-, MCP-1, MIP-1
Neutrophil elastase Neutrophil

Respiratory burst
Reactive oxygen species
IL-18
IL-3, IL-4, IL-10,
IL-13, IFN-, TNF-
Resting B cell
Resting T cell
IL-3, IL-6,
IL-7, GM-CSF
IL-3, IL-4,
IL-10
IL-10, IL-14,
IL-4, IL-6
Mast cell
Allergy anaphylaxis
IL-12, IL-15
Innate immunity
IL-3, IL-4, IL-5
Activated
LFA-1, VLA-4,
T cell
Mac-1
IFN-
Eotaxin
IL-2
IL-6
Activated B cell
Lung
epithelium
Eosinophil
IL-2
Viral infection,
tumor surveillance
IL-23
NK cell
TGF-, IL-2,
IL-4,IL-5, IL-13,
IFN-
Plasma cell
Th17/22
Th1/Th2
Th9
IgE
IgM/G/A
Allergy anaphylaxis
Humoral immunity
IL-2, IFN-, IL-4

Viral infection,
tumor surveillance
CTL
Fibrosis
IL-17A, IL-17F,
IL-22, IL-6,
GM-CSF
Autoimmunity
Tissue repair,
protozoan immunity
The inflammatory response in immunology.
15
Cancer research
Cancer research
At Life Technologies, we are dedicated to advancing cancer research and the

pursuit of a world without cancera goal we share with you, the research
and medical community. We are here to provide you with the instruments,
reagents, and technologies for cancer genomic and cancer cell analysis
applications to help translate your ideas from basic cancer research to
future clinical applications.
Translating genomic discoveries from the bench to the bedside and beyond
The key to fighting cancer through better therapeutics depends on a better understanding of the basic biology of this disease.
A major challenge in cancer research is unraveling the causes of cancer on a molecular level. Because most of the genomic
mutations or alterations in cancer impact the way that cells communicate with other cells, a solid understanding of both
genomic and cellular mechanisms is needed to accelerate cancer research.
At Life Technologies, we are committed to providing the widest technology selection with the highest quality at every budget to
help address challenges in cancer genomics and cancer cellular analysis applications.
Cancer Cellular Analysis Applications
Cancer Genomics Applications

Targeted
Sequencing
& Gene Panels
Cell Viability &

Proliferation
Cancer Cell
Imaging
Cell Signaling
& Pathway
Analysis
Cancer Cell
Growth &
Manipulation
Cancer
Stem Cells
Epigenetics
Cancer
Immunotherapy
Observe, follow, and probe deeper into cellular networks
Whole Genome
& Whole Exome
Sequencing
Gene Expression
by Sequencing
Genotyping
Assays-Somatic
Germline
Gene ExpressionFocused panels

& assays
Whole
Transcriptome
Identify changes in the genomes of tumors

that drive cancer progression
Overview of key cancer cellular analysis and genomics applications.
16

Cancer cellular analysis and genomic applications
Cancer cellular analysis involves understanding complex defects in communication between cells that alter
normal programs of proliferation, transcription, growth, migration, differentiation, and death.
Cancer research
Further advances in understanding altered cancer pathways and their interconnections may accelerate the
development of targeted molecular therapies allowing improvements in cancer diagnosis and treatment.
The right mix of tools and technologies are needed to observe, follow, and probe deeper into cancer cellular networks. With expertise that spans from cell culture, RNAi, and gene expression analysis to innovative products for rapid protein detection and cellular
imaging, we have affordable, high-quality solutions designed to help you reach your cancer research goals.
Learn more about the tools we offer for these applications below.
Cell viability, proliferation, and function
Evaluate the hallmarks of cancer with
Molecular Probes assays validated on
multiple instrument platforms, including
microscopes, flow cytometers, microplate readers, and high-content screening
instruments. To learn more, visit lifetechnologies.com/cellviability.
Cellular imaging
Visualize intracellular changes in cancer
cells right on your benchtop using Molecular Probes assays and the FLoid Cell
Imaging Station. To learn more, visit
lifetechnologies.com/floid.
Image cell cycle progression in live cells
with the Premo FUCCI Cell Cycle Sensor.
To learn more, visit lifetechnologies.com/
premochameleon.
Cell signaling pathways
Analyze how cancer cell signaling pathways are disrupted in cancer by accurate
quantitation of intracellular and extracellular proteins using protein analysis tools
designed for single or multiplex cancer
biomarker analysis. To learn more, visit
lifetechnologies.com/immunoassay and
lifetechnologies.com/abfinity.
Cell growth and manipulation
Foster the right cells and culture
environments to decipher the cancer
cell signaling pathways influenced
by altered genes. To learn more, visit
lifetechnologies.com/cellculture.
Identify genes and processes regulated
by your gene or pathway of interest
through the power of RNAi. To learn
more, visit lifetechnologies.com/rnai.
Use targeted editing of the genome to
precisely evaluate cancer phenotypes
through target validation and cellline optimization. To learn more, visit
lifetechnologies.com/geneart.
Rare cell event analysis

Identify, track, and analyze cancer cell
signaling events in individual cancer cells
using flow cytometry to study cell proliferation, the cell cycle, apoptosis, and immunophenotyping and to detect rare cellular
events such as the presence of circulating
endothelial cells. To learn more, visit
lifetechnologies.com/attune.
Cancer stem cells
Identify quiescent cancer stem cells and
determine their role in metastasis and
therapeutic resistance. To learn more,
visit lifetechnologies.com/cancerstemcells.
Cancer immunotherapy
Efficiently isolate, activate, and culture
various types of immune cells for your
cancer immunotherapy research needs.
immunotherapyresearch.
Drug discovery and development
Drive efficiency in drug assay development,
high-throughput screening, and metabolic
and safety assessment with the support,
expertise, and customization to meet your
unique cancer research needs. To learn more,
visit lifetechnologies.com/drugdiscovery.
Genotype-to-phenotype research
Determine normal and cancer-related functions of genes, and the implication of variation within those genes, to reach a better
understanding of cancer. To learn more, visit
lifetechnologies.com/cancergenotype.
Cancer epigenetics
The initiation and progression of cancer,
traditionally seen as a genetic disease, is
now realized to involve epigenetic abnormalities along with genetic alterations.
Recent advancements in the rapidly
evolving field of cancer epigenetics have
shown that disruption of various epigenetic mechanisms, including noncoding
RNAs such as microRNAs, DNA methylation, histone modifications, and nucleosome positioning, are central to switching
normal cells to malignant cells.
Life Technologies has developed powerful
epigenetic research technologies spanning renewable antibody development,
sample preparation, sequencing, quantitative PCR, and functional analysis assays.
cancerepigenetics.
Renewable epigenetic antibodies
Epigenetic regulatory proteins are involved
in a wide variety of chronic diseases,
including cancer, and are the target of an
explosive new field of drug discovery. The
ability to properly study most epigenetic
changes, which determine the genes that
are turned off and on, are determined by
the effectiveness of one critical toolantibodies. Life Technologies has partnered
with the Structural Genomics Consortium
to develop high-quality recombinant antibodies to read, write, and erase epigenetic
code. To learn more, visit lifetechnologies
.com/epiantibodies.
Noncoding RNAs that regulate cancer
Discovering how noncoding RNAs regulate gene expression will lead to new
approaches to understanding cancer.
At Life Technologies, we are developing sensitive methods to uncover new
RNAs, solutions to profile and validate
microRNAs, and functional analysis tools
to help you define the most important
noncoding RNAs in cancer. To learn more,
visit lifetechnologies.com/cancerrna.
Cancer genomics
We are committed to accelerating
cancer research with highly accurate
and sensitive sequencing and real-time
PCR systems to quickly find the cancer
mutations that matter. To learn more, visit
lifetechnologies.com/cancerbiomarkers.
17
Stem cell research

Stem cell research
The discovery in 2006 that human and mouse fibroblasts could be reprogrammed
to generate induced pluripotent stem cells (iPSCs) with qualities remarkably
similar to embryonic stem cells has created a valuable new source of pluripotent
cells for drug discovery, cell therapy, and basic research.
Life Technologies products have been an integral part of stem cell research from
before the discovery of iPSCs to current breakthroughs. With Gibco stem cell
culture products, Invitrogen stem cell reprogramming products, and a wide array
of stem cell analysis tools that include Molecular Probes labeling and detection
reagents, we have the products to support you at every step of your research.
Stem cell research can be time-consuming

finding support shouldnt be
Life Technologies gives you more PSC resources than any other sourceeasy
access to more than 70 proven protocols, >2,500 peer-reviewed publications, how-to videos, and webinars from leaders in the
stem cell community. And when you have a question, technical support is always available by phone, email, live chat, or in person.
We offer resources for areas of stem cell research from culture to reprogramming and tools for stem cell analysis and detection.
Culture4 media options, >20 protocols, and 50 years of innovation

Reprogramming3 reprogramming tools, 9 protocols, 4 webinars, and scientists standing by
Detection2 iPSC imaging methods, 1 instrument, and >30,000 colleagues
For more information about our resources for stem cell research, go to lifetechnologies.com/pscresources.
Culture
Whether youre studying stem cells for basic research, drug discovery, or therapeutic applications, Life Technologies offers an
extensive range of Gibco media, supplements, and reagents to meet all of your stem cell culture needs. You can culture and
expand stem cells in cGMP-manufactured serum-free, feeder-free, or xeno-free cell culture systems. Manufactured with superior
quality standards, our products offer the consistency and accuracy you need.
Find out about all of the options for PSC culture at lifetechnologies.com/culturepsc.
Reprogramming
Selecting the right reprogramming tool to generate iPSCs can leave you with questions. And no one has more answers
than Life Technologies. From traditional lentivirus methods to nonintegrating technologies like the CytoTune-iPS Sendai
Reprogramming Kit and episomal iPSC reprogramming vectors, our products provide more solutions to meet your safety, efficiency,
and budgetary needs.
Find out about all of the options for reprogramming at lifetechnologies.com/cellreprogram.
Detection
Validation is critical for iPSC research. Whether you want to quickly confirm that cells are pluripotent or need flexibility in antibody
and dye choice, Molecular Probes labeling and detection assays and imaging stations help you get the confirmation you need.
And, all of our labeling and detection technologies are backed by our extensive internal and external support resources, including
the Molecular Probes online community and the trusted Molecular Probes Handbook.
Find out about all of the options for detection at lifetechnologies.com/detectpsc.
18

Culture systems
KnockOut Serum Replacementwell-referenced supplement for
feeder-dependent growth of PSCs
Stem cell research
StemPro hESC SFMmedium for feeder-free growth of human PSCs

that allows optimization of basic fibroblast growth factor concentration
Essential 8 Mediumoptimized 8-component medium for feeder-free

growth of human PSCs
KnockOut SR XenoFree Kitfeeder-free and xeno-free medium for

human PSCs, with documentation for cell therapy applications
Reprogramming systems
Lentiviral particlesflexible technology for standard reprogramming

lifetechnologies.com/lenti
Episomal iPSC reprogramming vectorsnonintegrating, nonviral

technology for iPSC generation
CytoTune-iPS Sendai Reprogramming Kitnonintegrating technology

for standard and difficult cell types
Detection systems
Alkaline Phosphatase Live Stainallows for a quick check of pluripotency without compromising cell integrity
Antibodiesoptions for specific staining and visualization of stem cell

markers with maximum flexibility
FLoid Cell Imaging Stationbenchtop instrument for easy visualization

of Alkaline Phosphatase Live Stain and antibodies
19

Understanding the structural and functional relationships of plant cells and tissues is critical to plant research.
From studying the morphology of plant cells and walls, organelles, and other cellular compartments all the way
to complex gene regulation and expression studies, cell tissue analysis techniques and tools are critical not only
for basic plant research but also for programs of crop improvement. For example, demonstrating the mechanism of action of a new trait as part of the regulatory package is critical in trait development of new commercial
plant varieties. Key techniques such as immunohistochemistry have been routinely used to label diverse plant
tissues and organelles including cell walls, vacuoles, chromatin, nuclei, nuclear membranes, and chloroplasts.
To empower your research in plant cell and tissue analysis, Life Technologies has a broad portfolio of fluorescent
reagents, kits, and instruments specifically designed with your needs in mind.
Molecular Probes fluorescent dyes and probes
Molecular Probes reagents including Alexa Fluor dyes, Qdot nanocrystals, Click-iT detection assays, and
ProLong Gold antifade reagent are used in a wide range of plants, for a variety of plant research applications.
For example:
20
Immunohistochemistry
Fluorescence in situ hybridization
Organelle and cytoskeletal stains
Endocytosis studies
Transport studies
Glucose metabolism
Viability staining
Glutathione detection
Calcium and other ion imaging
Rubisco localization in maize leaf section. Rubisco was

localized using a rabbit anti-rubisco antibody and visualized using the highly cross-adsorbed Alexa Fluor
488 goat antirabbit IgG antibody (Cat. No. A11034).
The 2.0 m maize leaf section illustrates the immunolocalization of rubisco in the chloroplasts of the bundle
sheath cells surrounding the vascular bundles. The
red fluorescence, localized to the mesophyll plastids,
is due to background autofluorescence of chlorophyll.
Lignin appears dull green and is localized to the xylem
of the vascular bundle; cutin appears bright green and
is localized to the cuticle outside the epidermis. Image
contributed by Todd Jones, DuPont.
Pectin associated with plasmodesmatal pit fields of

kiwifruit cells. Pectin, a component of the cell wall
matrix and the main constituent of the middle lamella
that forms between daughter cell walls, was tagged
with an anti-pectin monoclonal antibody, JIM 5. The
primary antibody was detected and visualized with
Alexa Fluor 488 goat antirat IgG (Cat. No. A11006).
The primary antibody was a gift from Dr. Paul Knox,
University of Leeds, UK. Image contributed by Paul
Sutherland, The Horticulture and Food Research Institute of New Zealand, Ltd., Mt. Albert Research Centre.
Ca2+ gradient in elongating lily pollen tube. Top panel:
Pseudocolored image of a pollen tube of Lilium longiflorum injected with fura dextran (Cat. No. F3029).
The cell continues elongating and clearly shows a
Ca2+ gradient. Bottom panel: The same pollen tube
after injection with dibromo-BAPTA (Cat. No. D1211)
remains healthy but is no longer elongating. Images
contributed by Debra Miller, Dale Callaham, David
Gross, and Peter Hepler, University of Massachusetts.

Flow cytometry
Flow cytometry represents an ideal means for the analysis of both cells and subcellular
particles, with a potentially large number of parameters analyzed rapidly, simultaneously,
and quantitatively, thereby furnishing statistically exploitable data and allowing accurate
and facilitated detection of subpopulations. Since the first report of the use of this technique in plants 40 years ago, flow cytometry has become an essential tool for the understanding of fundamental mechanisms and processes underlying plant growth, development, and function. Key applications include the study of ploidy levels, genome mapping,
localization of sex-specific chromosomes, and plant cell and algal studies.
Life Technologies offers a wide range of flow cytometric analysis tools including the
Attune Acoustic Focusing Cytometer and fluorescent probes.
Learn more and see all of our products at lifetechnologies.com/probes.
Featured product: Alexa Fluor dyes
The leading and most trusted fluorescent dyes available today have been used extensively
in the plant sciences. In addition to superior performance, youll get an experienced,
problem-solving technical support team, over 30,000 published references, application
and experimental tips, and protocols to help with experimental planning. Our range of
labeling kits, molecules for tracing cell structures, and secondary detection reagents
within the Alexa Fluor product line provide superior brightness and photostability,
outperforming conventional fluorescent reagents.
Learn more at lifetechnologies.com/alexa.
For a comprehensive view of all your products and technologies to empower your plant
sciences research, visit lifetechnologies.com/plants.
106
SSC-A
106
PI 640 LP BL3-A
A
Scatter
105
104
103
103
104
105
105
Nuclei
104
103
104
106
105
PI 603-48 BL2-A
700
1,000
900
4C
600
2C
8C
600
500
400
300
200
200
100
0
104
4C
700
Count
Count
300
2C
800
500
400
106
PI 603-48 BL2-A
16C
32C
105
PI 603-48 BL2-A
8C
100
105
16C
50
100
150
200
250
300
PI 603-48 BL2-A (10^3)
Nuclear holoploid genome sizing analysis of Arabidopsis thaliana leaf tissue homogenates using the Attune Acoustic
Focusing Cytometer. (A) Biparametric density plot of side scatter vs. propidium iodide (PI) fluorescence with Scatter
gate surrounding the fluorescent nuclei. (B) Biparametric density plot of PI fluorescence (603/48 vs. 640 nm) of
Scatter gate population from A. (C) Logarithmic histogram of PI fluorescence (603/48 nm) of Nuclei gate population
from B. (D) Linear histogram of PI fluorescence (603/48 nm) of Nuclei gate population from B. 2C, 4C, 8C, 16C, and
32C denote the C-value for the respective peaks.
21
AKT pathway
AKT pathway
The serine-threonine kinase AKT (also known as protein kinase B) is a protein that acts as a central convergence node in a broadly influential signaling network. AKT activation serves as a master switch of these
cellular signaling pathways, generating a multitude of intracellular responses through a plethora of downstream targets and interacting partners. Because of its pivotal role in cell signaling, and the consequences
of that signaling in diseases ranging from cancer and diabetes to neurodegeneration, AKT is one of the most
actively studied kinases in both basic research and drug development. Here we highlight a few of our products for analyzing the AKT signaling pathway and its downstream targets.
Life Technologies offers Novex antibodies, ELISAs, Novex assays for the Luminex platform, and growth
factors for key targets in the AKT signaling cascade.
Key targets of interest in the AKT pathway

Protein
Characteristics
Means of regulation
Research tools (Chapter 4, pages 146-155)
AKT/protein kinase B
Master regulator of cell growth and

signaling cascade
Phosphorylation/
dephosphorylation
Novex antibodies, ELISAs, and Novex assays

for the Luminex platform
mTOR
Cell growth regulator
Phosphorylation/
dephosphorylation

GSK-3B
Cell cycle regulator
Phosphorylation/
dephosphorylation

MDM2
Negative regulator of p53 function
Phosphorylation/
dephosphorylation

NF-B
Transcriptional activator regulating

apoptosis
Phosphorylation/
dephosphorylation

Additional references
Madonna R, Bolli R, Rokosh G et al. (2012) Targeting phosphatidylinositol 3-kinase-Akt through hepatocyte growth factor for
cardioprotection. J Cardiovasc Med doi: 10.2459/JCM.0b013e3283542017.
Papadimitrakopoulou V (2012) Development of PI3K/AKT/mTOR pathway inhibitors and their application in personalized therapy
for non-small-cell lung cancer. J Thorac Oncol doi: 10.1097/JTO.0b013e31825493eb.
Wang G, Pan J, Chen SD (2012) Kinases and kinase signaling pathways: potential therapeutic targets in Parkinsons disease. Prog
Neurobiol 98(2):207221.
Sheppard K, Kinross KM, Solomon B et al. (2012) Targeting PI3 kinase/AKT/mTOR signaling in cancer. Crit Rev Oncog 17(1):6995.
22
Growth factor
PDK1
mTORC2
mTOR
GL
+P
AKT
+P
Cytoskeletal
reorganization
+P
PP2A
4
PRAS40
Rheb
GL
mTOR
Cell growth
raptor
Receptor tyrosine kinase
Hormone receptor
+P
eIF4B
MDM2
CASP9
FOXO
Cell cycle
+P
IB
-CTN
NFAT
IkB
NFB
eIF2B
p53
Glycogen
synthesis
Translation
initiation
Apoptosis
IKK
+P
RPS6
Phosphatase
BAD
CCND1
+P
Kinase
AR
+P
+P
eIF4A
Translation factor
+P
GSK-3
+P
eIF4E
Transcription factor
+P
+P
PDCD4
ASK-1
p27
SK6
4E-BP1
PI3K
+P
p21
+P
+P
+P
+P
+P
mTORC1
FKBP
+P
RTK
+P
+P
-P
+P
Rapamycin
PTEN
p85 p110
+P
rictor
TSC2
TSC1
PIP2
-P
AKT pathway
PIP3
FOXO
GATA4 NFAT NFB
Transcription
FAS-L
p27
KIP1
p21
FAS
PTEN
CIP1
Cell proliferation
The AKT pathway.
23
MAPK pathway
MAPK pathway
The mitogen activated protein kinase (MAPK) pathway is central to cellular signal transduction in response
to growth factors, mitogens, cytokines, and stress. The MAPK pathway is regulated through a series of
reversible phosphorylation events resulting in a variety of cellular activities including growth, differentiation, development, and apoptosis. Here we highlight a few of our products for analyzing the MAPK signaling
pathway and its downstream targets.
factors for key targets in the MAPK signaling cascade.
Key targets of interest in the MAPK pathway

Protein
Characteristics
Means of regulation
MAPK
Regulator of cell growth and signaling

cascade
Phosphorylation/
dephosphorylation
Novex antibodies, ELISAs, and Novex assays for

the Luminex platform
ERK
Regulator of growth, development, and

poliferation
Phosphorylation/
dephosphorylation

JNK
Regulator of inflammation, apoptosis,

and differentiation
Phosphorylation/
dephosphorylation

Ras
GTPase involved in cell proliferation
Cleavage of GTP

Oeztuerk-Winder F, Ventura JJ (2012) The many faces of p38 mitogen-activated protein kinase in progenitor/stem cell differentiation. Biochem J 445(1):110.
Gantke T, Sriskantharajah S, Sadowski M et al. (2012) IB kinase regulation of the TPL-2/ERK MAPK pathway. Immunol Rev
246(1):168182.
Ppulo H et al. (2012) The mTOR signalling pathway in human cancer. Int J Mol Sci 13(2):18861918.
Aksamitiene E, Kiyatkin A, Kholodenko BN (2012) Cross-talk between mitogenic Ras/MAPK and survival PI3K/Akt pathways: a fine
balance. Biochem Soc Trans 40(1):139146.
Davies C, Tournier C (2012) Exploring the function of the JNK (c-Jun N-terminal kinase) signalling pathway in physiological and
pathological processes to design novel therapeutic strategies. Biochem Soc Trans 40:8589.
24
MAPK pathway
The MAPK pathway.
25
JAK/STAT pathway
JAK/STAT pathway
The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway plays a central role in
transducing signals from a wide array of cytokines and growth factors leading to various cellular functions,
including proliferation, growth, development, hematopoiesis, and immune responses.
Abnormal constitutive activation of the JAK/STAT pathway has been implicated in various cancers and immune
disorders. Thus, inhibitors of the JAK/STAT pathway are being sought.
Here we highlight a few of our products for analyzing the JAK/STAT signaling pathway and its downstream targets.
factors for key targets in the JAK/STAT signaling cascade.
Key targets of interest in the JAK/STAT pathway

Protein
Characteristics
Means of regulation
JAKs
Initiate signaling upon receptor

multimerization
Phosphorylation/dephosphorylation Novex antibodies, ELISAs, and Novex assays

STATs
Transcription factors
Phosphorylation/dephosphorylation,
DNA binding

SOCS
Regulator of JAK signaling
Binding of JAK

PIAS
Regulator of STAT signaling
Binding of STAT

Harrison DA (2012) The Jak/STAT pathway. Cold Spring Harb Perspect Biol 4(3). pii: a011205. doi: 10.1101/cshperspect.a011205.
Sansone P, Bromberg J (2012) Targeting the interleukin-6/Jak/stat pathway in human malignancies. J Clin Oncol 30(9):10051014.
Wagner KU and Schmidt JW (2011) The two faces of Janus kinases and their respective STATs in mammary gland development and
cancer. J Carcinog 10:32. doi: 10.4103/1477-3163.90677.
Mohr A, Chatain N, Domoszlai T et al. (2012) Dynamics and non-canonical aspects of JAK/STAT signalling. Eur J Cell Biol 91(67):524532.
Tamiya T, Kashiwagi I, Takahashi R et al. (2011) Suppressors of cytokine signaling (SOCS) proteins and JAK/STAT pathways: regulation
of T-cell inflammation by SOCS1 and SOCS3. Arterioscler Thromb Vasc Biol 31(5):980985.
26
JAK/STAT pathway
The JAK/STAT pathway.
27
Neurodegeneration
Neurodegeneration
Neuronal death is pivotal to Parkinsons disease (PD) pathogenesis and can be triggered by various cellular
mechanisms, including oxidative stress, misfolding of proteins, mitochondrial dysfunction, and proteasome
dysfunction. In their pursuit to fully understand the various pathways that lead to neurodegenerative diseases
such as PD, researchers are exploring the molecular basis of these mechanisms. Here we highlight just a
few of our products for analyzing the key factors and effectors in PD.
Life Technologies offers Novex antibodies, ELISAs, Novex assays for the Luminex multiplex assays
platform, and growth factors for key targets in studying Parkinsons disease.
Some key targets of interest are listed in the table below.
Protein
Characteristics
Means of regulation
Parkin
Mitochondrial health
Mutation and misfolding
Novex antibodies
-Synuclein
Mitochondrial health
Aggregation and mutation
Novex antibodies, ELISA, and Novex assays for

LRRK2
Leucine-rich kinase
Phosphorylation/dephosphorylation

Tau
Nucleation and stabilization

of microtubules
Phosphorylation/dephosphorylation/
misfolding

Protter D, Lang C, Cooper AA (2012) Synuclein and mitochondrial dysfunction: a pathogenic partnership in Parkinsons disease?
Parkinsons Dis 2012:829207.
Sutachan JJ, Casas Z, Albarracin SL et al. (2012) Cellular and molecular mechanisms of antioxidants in Parkinsons disease. Nutr
Neurosci 15(3):120126.
Harvey L, Boksa P (2012) Prenatal and postnatal animal models of immune activation: Relevance to a range of neurodevelopmental disorders. Developmental Neurobiology special issue: Neuroimmunology in Development and Disease. Dev Neurobiol doi:
10.1002/dneu.22043.
Wang G, Pan J, Chen SD (2012) Kinases and kinase signaling pathways: potential therapeutic targets in Parkinsons disease. Prog
Neurobiol 98(2):207221.
Rademakers R, Neumann M, Mackenzie IR (2012) Advances in understanding the molecular basis of frontotemporal dementia.
Nat Rev Neurol doi: 10.1038/nrneurol.2012.117.
28
Neurodegeneration
Neurodegeneration.
29

Delivering speed, scalability, and quality
Scalability is critical for clinical and biomanufacturing applications. Our tech transfer process helps ensure smooth scale-up as
batch size increases.
Small-scale media production when speed

matters most
Large-scale media production in our worldclass, validated manufacturing facilities
Choose Gibco MediaExpress for small-scale media

production:
Choose Gibco Custom Cell Culture Services for large-scale

media production:
Fast turnaround (typically <10 days)

Small-scale batches (<200 L, <10 kg)
Non-cGMP production
5,000 ft2 pilot plant, Grand Island, NY
Industry-leading quality systems and support

Large-scale batches (up to 10,000 L)
Learn more at lifetechnologies.com/custommedia.
Learn more at lifetechnologies.com/mediaexpress.
30

Gibco PD-Direct Bioprocess Services provides media development and
optimization services
Gibco PD-Direct services provide contract services leveraging the latest technologies and products from
Life Technologies to solve cell-line, media, and process development challenges. PD-Direct services can be
your reliable outsourcing partner to expand your capabilities and deliver on-time exceptional results.
Choose Gibco PD-Direct Bioprocess Services to partner with technical and industry experts who can
assist you every step of the way. Learn more at lifetechnologies.com/bioproduction.
Customize the media format, quality control (QC) testing, and documentation to your needs
Choose a media format and batch size:

Liquid: 1 to 10,000 L
Powder: 1 kg to 8,000 kg
Concentrates: 10 to 5,000 L
Gibco AGT media: 2 kg to 6,000 kg
Select packaging based on workflow:
Custom, or off-the-shelf for reduced lead time
Closed system, ideal for aseptic processes and applications
Solutions for disposable workflows
Select QC testing, documentation:
Standard QC testing: pH, osmolality, and sterility
Documentation: Certificate of Analysis (COA), Material Safety Data Sheet (MSDS),
import/export documentation
Additional testing and documentation available upon request
Which custom media solution is best for you?

Gibco MediaExpressSM Services
Gibco Custom Cell Culture services
Gibco PD-Direct Bioprocess

Services
Description
Catalog modification or users

formulation
Catalog modification or users

formulation
Media development or
optimization projects
Regulatory
Non-cGMP
cGMP
Non-cGMP
Batch sizes
Liquid: 1200 L
Powder: 110 kg
AGT media: 210 kg
Liquid: 1010,000 L
Powder: 18,000 kg
AGT: 506,000 kg
Based on scope of work (SOW)
Lead time
2 weeks
1216 weeks
Variable
Packaging sizes
Liquid: 100 mL, 500 mL, 1 L bottles; 5 L,

10 L bags
Powder/AGT media: 110 kg; range of
plastic containers
Liquid: 100 mL, 500 mL, 1 L bottles;

1200 L range of bags
Powder/AGT media: 110 kg; range of
plastic containers; 55 gal HDPE drums
Liquid: 100 mL, 500 mL,

1 L bottles; 5 L, 10 L bags
Powder/AGT media: 110 kg;
range of plastic containers
Testing
Check and record
Standard QA/QC release
Customizable
Documentation
Formulation instructions
Certificate of Analysis (COA)
Based on scope of work (SOW)
For more information
lifetechnologies.com/custommedia
lifetechnologies.com/custommedia
Email: pd_direct@lifetech.com
To get started, request a quote for custom media now.

Customize a Gibco catalog media product at lifetechnologies.com/mediaconfigurator.
Have us manufacture your own custom media formulation at lifetechnologies.com/custommedia.
31
Custom Novex multiplex assays for

Luminex technology
New made-to-order assay design system: design and place an order or

get a quote online
Custom Novex multiplex assays for Luminex technology
Our custom Novex multiplex immunoassay kits for the Luminex platform maximize flexibility in experimental design, permitting the quantitation of one or multiple targeted proteins in unique panels designed by you. For your convenience, each customized panel comes with reagents that are blended, optimized, tested, and designed for use with the Luminex 100/200 system,
FLEXMAP 3D system, and the new MAGPIX system.
Choose specific targets of interest

(polystyrene or magnetic)
Combined with appropriate

Buffer Kit (all needed reagents)
for ease of use
Each assay is premixed and QC tested

for optimum top signal, low
background, and linearity of dilution
Target
Species
Cytokine
Human
Magnetic
Chemokine
Mouse
Polystyrene
Growth factor
Rat
Signaling protein
Nonhuman primate
Bead type
Design and price your own Novex multiplex assay kit at bioinfo.invitrogen.com/luminex.
32
Drug discovery demands custom research and custom solutions

From cell engineering to custom assay development, our scientists work closely with your team to understand your goals and tailor
a solution that fits your unique project guidelines.
When an off-the-shelf tool isnt the right option, our custom biology group offers unique and highly efficient solutions to help ensure
you stay on track. Expect consistently short cycle times and high-quality results on time, every time, with proactive communication
throughout the project.
Custom assays and services
Custom biochemical and cell-based assaysLet Life Technologies create a
custom biochemical or cell-based solution to suit your needs, using the broad
array of technologies in the Life Technologies toolbox.
Cell line generationOutsource your cell line development and let us craft a
Jump-In parental cell line or final engineered cell line that meets your defined
specifications.
Antibody labelingChoose from europium or terbium donor chelate labels and
obtain labeled antibodies for your TR-FRET assay development projects.
Cell provisioningGet validated, high-throughput screeningready cells in large
scale with our cryopreservation service.
siRNA profilingIdentify and validate your target of interest using first-in-class
siRNA collections with readouts to interrogate functional pathways, proliferation,
gene expression analysis, and protein modifications.
BacMam virus production assaysLet us create a BacMam virus from your gene
of interest or develop a portable and robust BacMam-enabled assay.
TaqMan protein assaysGet highly sensitive protein detection coupling the specificity of antibody affinity reagents with the sensitivity and fidelity of TaqMan PCR.
Tell us your custom assay requirements at lifetechnologies.com/custom.
Biochemical and cellular profiling services
Drug discovery scientists have trusted the SelectScreen Services team for thousands
of screening and profiling projects on more than 500 targets.
SelectScreen Services are executed with strict quality controls using a state-of-theart compound management system to drive down cycle times and meet expanding
customer demand. Highly trained personnel and automated processes driven by integrated informatics systems help us ensure that reliable, high-quality data are delivered every time.
SelectScreen Kinase Profiling Serviceprofile with our expanding panel of

distinct protein kinases and get an average delivery time of just 6 days.
SelectScreen Cell-Based Pathway Profiling Serviceunderstand potential

off-target effects earlier with our rapid, flexible cell-based profiling service against
a panel of pathway-specific CellSensor cell lines.
SelectScreen Cell-Based GPCR Profiling Servicechoose from a panel of
functionally validated GeneBLAzer, second-messenger, and Tango betaarrestin recruitment cell-based assays.
SelectScreen Cell-Based Nuclear Receptor Profiling Serviceprofile your
compounds against the most comprehensive panel of target-specific GeneBLAzer nuclear hormone receptor assays for early safety and liability testing.
Choose the right profiling service for your research at lifetechnologies.com/selectscreen.
33
Instructional videos and companion handbook introduce the

fundamentals of cell culture
The Gibco Cell Culture Basics videos and handbook provide critical training and troubleshooting for each essential cell culturing
technique. Following these protocols will help your lab attain reproducible results every day. The handbook is an introduction to cell
culture, covering the key topics listed below:
Cell culture laboratory safety

Cell culture equipment
Aseptic technique
Aseptic technique checklist
Biological contaminationtypes, cross-contamination,
and use of antibiotics
Cell linesacquiring & selecting

Adherent cell culture vs. suspension cell culture
Cell culture environment (media, pH, CO2, and
temperature)
Mammalian and insect cell morphology

Cell culture protocols
Guidelines for maintaining cultured cells
Subculturing adherent cells
Subculturing suspension cells
Freezing cells
Thawing frozen cells
Counting cells in a hemocytometer
Trypan blue exclusion
Cell culture troubleshooting
Cell culture products
Transfection and selection
The video series includes all basic cell culture techniques

through live demonstration. The video titles include:
Introduction to Cell Culture

Sterile Technique
Passaging Cells
Freezing Cells
Thawing Cells
Learn more about these videos and protocols at lifetechnologies.com/cellculturebasics.
Cell Line Database

The Cell Line Database is a Web-based tool that allows researchers
to find more information about cell lines theyre already using,
to find new cell lines, or to find compatible Life Technologies
products and applications. The listing is alphanumeric and
provides a rapid source of information about the type of cell,
species, source type, and behavior of the cell in applications.
The Cell Line Database can be found at lifetechnologies
.com/celllinedatabase.
Example search for HeLa cells

Technical ResourcesCell Line Database
Cell Line Detail
Cell Line Name: HeLa cells
Organism Type:
Tissues:
Cervix
Adherence Phenotype: Adherent
Primary:
No
Scientific
application(s):
Transient Transfection
Products and applications for HeLa cells

Gene Expression 7
Media/Cell Culture 15
Nucleic Acid Purification 1
Selective Antibiotics 1
Transfection Display 23
= Protocol exists within search results
34
Human
Protocol Database
The Protocol Database is a Web-based tool that allows researchers to find more than 500 validated protocols for performing critical cell biology experiments. Protocols include culturing primary or stem cells,
performing imaging and other cell analysis techniques, and protein purification, separation and identification techniques. The database allows the user to search for a specific protocol using keywords or to browse.
The database can be found at lifetechnologies.com/protocolsdatabase.
Protocol Database
The Molecular Probes

Handbook online
The online version of The Molecular Probes Handbook (lifetechnologies.com/handbook) is a comprehensive
resource for fluorescence technology that is regularly updated. Like the printed version, the online Handbook
contains detailed information about the thousands of Molecular Probes products available for fluorescence
and imaging applications. The online Handbook has product pricing for your region as well as the added
advantage of having critical product information at your fingertips. Find easy access to data images and
referenced citations as well as product structures and spectra.
35
Fluorescence
SpectraViewer
The Fluorescence SpectraViewer is an online tool that can be

used to plot and compare the excitation and emission spectra
of fluorescent dyes, facilitating the design of multicolor fluorescence experiments. As many as five fluorophores can be
selected from an extensive list of dyes for simultaneous spectral analysis to determine the suitability of fluorophores for
specific instrument configurations, and to evaluate spectral
compatibility of different fluorophores. The SpectraViewer is
available at lifetechnologies.com/spectraviewer.
Mobile applications
and widgets
Life Technologies offers an assortment of handy applications
and widgets for your everyday mobile needs. Download popular
iPhone apps and widgets from Molecular Probes and other
Life Technologies brands, including the DailyCalcs Science
Calculator and the Alexa Fluor Selection Guide widget. Visit
lifetechnologies.com/apps to get started. New apps and
widgets are introduced on a regular basis, so check back often.
Virtual Cell
Staining Tool
The Virtual Cell Staining Tool (lifetechnologies.com/
cellstainingtool) allows you to select different combinations of
cellular structures and fluorophores to create your perfectly
labeled fluorescent cell, which you can easily share with your
colleagues using our email or print functionality.
36
Antibody and immunoassay

selection guides
Product selection guide
Purpose
Immunoassay Selector
Tool
Find the right Novex immunoassay product easily using our convenient selection tool. Simply search by keyword,
then filter your results to find the right ELISA kit, Luminex platform assay, or antibody pair.
http://igene.invitrogen.com/products/selector/immunoassays
Labeling Chemistry
Selector Tool
Quickly find the best Molecular Probes reactive dye or label for your experiment. Whether you want to label
proteins and antibodies for immunofluorescence with our Alexa Fluor dyes, nucleic acids for in situ hybridization,
or lipids for membrane studies, this tool will help you find the right conjugation product.
http://igene.invitrogen.com/products/selector/dyes
Find all of the Life Technologies primary antibodies, including Molecular Probes flow cytometry antibodies,
Novex primary antibodies for western blot analysis, and more. Simply search by target, gene symbol, or gene ID,
then filter your results by application, reactivity, host, conjugate type, or antibody type. You can also search within
your results to quickly find the right antibody.
Antibody and immunoassay selection guides
Primary Antibody
Selector Tool
http://igene.invitrogen.com/isearch
Secondary Antibody
Selector Tool
Finding the right secondary antibody is easier than ever. Whether you are looking for secondary antibodies,
anti-dyes and anti-haptens, anti-epitope tags and anti-reporter genes, or isotype controls, Life Technologies has
the largest selection available. Simply select the type of antibody you are looking for and filter by target, Ig class,
reactivity, conjugate, or host.
http://igene.invitrogen.com/antibody
Customer and
technical support
Comprehensive worldwide support
Whether you need help with a current order, placing a new order, or finding more information about product availability, please contact your local customer service team.
Technical support and training
If you have questions about product selection or use, assay or experimental design,
data analysis, or troubleshooting, contact our team of technical support scientists or
utilize our comprehensive portfolio of online product and application support tools.
LSIA 2011 Awards Most Responsive

Customer Service & Most
Knowledgeable Technical Support
How to reach us
To find your local customer service or technical support team, go to lifetechnologies.com/contactus.

For product FAQs, protocols, training courses, and webinars, go to lifetechnologies.com/technicalresources.
Follow us on Twitter:
@LifeTechSupport
@AppliedBio
@dynabeads
@everydayprotein
@gibco
@invitrogen
@molprobes
@The_RNA_experts
@lifeimmunology
@LifetechEMEA
facebook.com/gibcocellculture
facebook.com/cellimaging
facebook.com/immunologynews
facebook.com/ambion
facebook.com/invitrogen
facebook.com/novexprotein
Like us on Facebook:
facebook.com/appliedbiosystems
facebook.com/lifetech.taqman
37
Gibco cell culture and cells

We have a 50-year heritage of bringing you dependable, consistent, and superior-quality cell culture products that provide scientists
like you with the means and the confidence to elevate your research. From cutting-edge stem cell research tools like Essential 8
Medium, the CytoTune-iPS Sendai Reprogramming Kit, and our broad collection of human primary cells, to everyday essential media
like DMEM and GlutaMAX Supplement, Gibco products help set the standard for cell culture in laboratories around the world.
Section
Highlighted
products
Primary and
stem cells
Primary human
keratinocytes,
fibroblasts, corneal
epithelial cells,
umbilical vein endothelial cells, and
more, page 56
Neural primary and
stem cells, page
56, 71
Mesenchymal stem
cells, page 68
Hematopoietic stem
cells, page 73
Recover
Grow
Recovery
Freezing Medium
results in 25%
more viable
cells post-thaw
and requires
no additional
supplementation,
page 40
Gibco FreeStyle protein expression systems provide rapid, scalable, highyield transient expression in both 293 and CHO cells, page 42
Gibco CD Hybridoma Medium is a chemically defined, animal originfree
medium optimized for the growth of hybridomas, page 42
Gibco Sf-900 III medium improves the consistency of your insect cell
culture, page 47
Choose Geltrex, AlgiMatrix, or CellStart CTS 3D cell culture matrices
to get more physiologically relevant culture conditions, page 49
Essential 8 Medium offers optimal feeder-free culture for all human PSC
and iPSC lines with minimal lot-to-lot variability, page 60
StemPro NSC SFM is a serum-free medium for neural stem cells, page
52, 63
EpiLife medium for enhanced primary keratinocyte cell culture, page 57
Cell Therapy Systems (CTS) validated reagents for clinical research applications, look for the CTS label throughout this chapter
Everyday
essentials
Classical Gibco media, cells, sera, and reagents for general cell culture
in a wide variety of formats and sizes. Choose from standard media such
as DMEM, RPMI, and MEM to more advanced formulations including
GlutaMAX, OptiMEM, or Advanced reduced-serum media, page 41
Gibco provides FBS for both general and specialized cell culture, page 43
Discover a broad range of eukaryotic and bacterial selection antibiotics such
as Geneticin, puromycin, and Zeocin, page 46
Gibco growth factors are manufactured for high activity, purity, and
compatibility with Gibco media, page 47
B-27 Supplement, the most published serum substitute for neural cell
culture, page 72
Neurobasal media, basal media formulated to sustain optimal culture of
neurons when used with B-27, N-2, or G-5 supplements, page 71
KnockOut Serum Replacement, the standard supplement for growing
pluripotent stem cells from multiple species, over 2,400 publications,
page 6061
Instruments
38
The chart below highlights our most popular products and newest technologies in cell culture. Follow the Web links provided in this
chapter for a complete listing of all our products and services.
If you have any questions about which of our solutions are right for you, please contact Life Technologies technical support by
phone, email, Facebook at facebook.com/gibcocellculture, or Twitter at twitter.com/gibco.
Passage
Reprogram/engineer
Detect
TrypLE Express, TrypLE Select, and

StemPro Accutase are gentle and effective
reagents for the dissociation of cell monolayers, page 52
The CytoTune-iPS Sendai Reprogramming

Kit generates iPSCs from challenging cell
types with this easy-to-use, nonintegrating
technology, page 64
Alkaline Phosphatase Live Stain is ideal for

detecting pluripotency, page 66
TrypLE Select 10X is ideal for strongly

adherent cells, page 52
Episomal iPSC reprogramming vectors are

integration-free and viral-free reprogramming vectors, page 64
Gibco liquid and powdered trypsin provide

fast and convenient general-purpose cell
dissociation, page 52
Countess Automated Cell Counter takes the

subjectivity and tedium out of counting live
and dead cells; fast, accurate, and convenient cell counting at your fingertips, page 53
39
Cell culture products

The Gibco brand of cell culture products has a 50-year heritage of dependability, consistency, and superior quality. Our goal is to
provide scientists like you the means and the confidence to elevate research and healthcare now and into the future. We sincerely
appreciate your trust, and are extremely proud to support your achievements thus farand the many yet to come. Explore our
entire catalog of cell culture products at lifetechnologies.com/cellculture.
Recover
Grow
Passage
Efficient and effective cryopreservation is

a critical step in cell culture maintenance.
Recovered samples that have a large number
of viable cells are the foundation of robust
and healthy cultures, and allow you to begin
experiments sooner and have more confidence in the results.
From the most basic formulations to the

newest innovations, Gibco products are
designed to provide the highest quality,
consistency, and performance for your cell
culture andtissue cultureneeds. These are
media, reagents, cells, sera, and supplements from the leader in cell culture.
Gibco cell dissociation products are ideal

for use with tissues and cell monolayers and
come in a wide variety of formats to meet
the diverse needs of researchers performing
adherent cell culture. Automated cell
counting improves the consistency of your cell
culture passage to passage.
Recover
Recovery Freezing Medium

Recovery Cell Culture Freezing Medium is a complete cryopreservation medium for
mammalian cell cultures.
Recovery medium results in an average of 25% increase in cell viability in
Cell culture
cryopreservation of both adherent and suspension cell lines
Recovery medium is an optimized, fully supplemented formulation which avoids

the messy mixing of DMSO
Recovery medium has proven performance on a broad range of mammalian

cell lines
Recovery medium contains DMEM, fetal bovine serum, calf serum, and 10%
DMSO. Recommended storage conditions: 5C to 20C
Learn more at lifetechnologies.com/recovery.
Grow
The following key cell culture interest areas are included in this section, with pages
listed.
Mammalian cell culture..............................4147
Insect cell culture........................................4748
Neurobiology media.....................................4849
Extracellular matrices.......................................49
Microbial culture................................................50
Cytogenetics.......................................................51
40
Mammalian cell culture

Classical media
We offer products to support the growth of a variety of different mammalian cell lines, including ready-to-use options
as well as powder and liquid formulations. Gibco cell culture media are time tested and cited in scientific literature
more than any other brand in the world.
Learn more about DMEM, RPMI Media 1640, MEM and other classical media at lifetechnologies.com/classicalmedia.
Gibco Dulbeccos Modified Eagle Medium (DMEM) is designed to deliver superior quality, consistency, and control for
your mammalian cell culture. We offer DMEM in a number of formulations.
Gibco RPMI Media 1640 performs well in a wide range of applications for mammalian cells, including the culture
of fresh human lymphocytes, fusion protocols, and growth of hybrid cells. We offer RPMI Media 1640 in a number of
formulations.
Gibco Minimal Essential Medium (MEM) is based on Eagles Media and is well suited for the growth of a broad spectrum of mammalian cells. We offer MEM in a number of formulations.
GlutaMAX media
Gibco GlutaMAX medium is our standard, trusted cell culture medium that contains a stabilized dipeptide form of
L-glutamine, L-alanyl-L-glutamine, which helps prevent degradation and ammonia build-up even during extended
culture periods.
Cell culture
Minimizes toxic ammonia build-up

Stable at room temperature
Improves cell viability and growth
Extremely stable in aqueous solution, the L-alanyl-L-glutamine dipeptide will not degrade into ammonia in storage
or incubation like L-glutamine. When cultured in medium that contains GlutaMAX-I supplement, cells gradually
release aminopeptidases that hydrolyze the dipeptide, which in turn slowly releases L-alanine and L-glutamine into
the culture media. The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production
or in the TCA cycle. This is analogous to the fed-batch culture strategy in which L-glutamine is continuously added to
the culture but maintained at a low concentration. The result is efficient metabolic processing and high growth yield
without excess ammonia.
We offer GlutaMAX-I supplement in DMEM, MEM, RPMI, Opti-MEM medium and others. You can also purchase the
GlutaMAX-I dipeptide and substitute it directly for L-glutamine in your current cell culture media formulation.
Learn more at lifetechnologies.com/glutamax.
Advanced media
Gibco Advanced reduced-serum media are enhanced basal media formulations of DMEM, DMEM/F-12, MEM,
and RPMI 1640. Enriched with normal-serum constituents, these media require 5090% less FBS supplementation,
deliver equivalent or superior cell growth compared to media supplemented with 10% FBS, and elicit no changes in
morphology or function in many common cell lines.
Learn more at lifetechnologies.com/advanced.
Opti-MEM media
Gibco Opti-MEM reduced-serum medium is an improved minimal essential medium (MEM) that allows for a reduction of FBS supplementation by at least 50% with no change to growth rate or morphology. Opti-MEM medium is also
recommended for use with cationic lipid transfection reagents, such as Lipofectamine reagent. Opti-MEM medium
can be used with a variety of suspension and adherent mammalian cells, including Sp2, AE-1, CHO, BHK-21, HEK,
and primary fibroblasts. Opti-MEM media are available in various formats for a range of cell culture applications,
including hybridoma culture and transient and stable protein expression cultures.
Learn more at lifetechnologies.com/optimem.
41
Hybridoma media
Improve serum-free hybridoma culture with Gibco media
Choose from chemically defined, protein-free, and serum-free formulations
that maximize performance
Animal originfree products help minimize risk

Custom products and packaging available
Easy to scale up to large-volume cultures
Learn more about hybridoma media at lifetechnologies.com/hybridoma.
Product
CD Hybridoma Medium*
(Cat. No. 11279023)
CD Hybridoma AGT
Medium* (Cat. No.
12372025)
Hybridoma-SFM*
(Cat. No. 12045084)
PFHM-II*
(Cat. No. 12040077)
Media type
Chemically defined medium
Chemically defined medium
Serum-free medium;
low-protein (20 g/mL)
Protein-free medium
Animal originfree
Yes
Yes
No
Yes
Optimized for
Human, mouse, rat hybridomas, myelomas. NS0, NS-1,

and other steroid-dependent cells when used with 250X
Cholesterol Lipid Concentrate (Cat. No. 12531018).
Applications
Growth and MAb production; can be used to express other proteins in engineered myeloma cell lines
Human, mouse, rat hybridomas, myelomas
Cell culture
* Drug Master File available.

Dry granular format of CD Hybridoma Medium.
Protein expression media

Mammalian transient expression systems enable large-scale production of proteins and are ideal for the
expression of human or other mammalian proteins because they generate recombinant proteins with closerto-native folding and post-translational modifications (i.e., glycosylation) than those expressed from E. coli,
yeast, or insect cells. Gibco optimized mammalian protein expression media for CHO and 293 suspension
cultures help you achieve rapid, scalable, high-yield protein production from transiently transfected cultures,
and offer easy scale-up and purification with serum-free and animal originfree components.
Learn more about the expression systems highlighted below at lifetechnologies.com/freestyle.
Expi293 Expression Systemhigh-density, high-yield,
scalable protein expression
FreeStyle MAX 293 Expression Systemnew and improved

FreeStyle MAX transfection reagent
Each system supplies:

FreeStyle 293-F cells
FreeStyle 293 Expression Medium
FreeStyle MAX Transfection Reagents
pCMVSPORT -gal vector
OptiPro SFM
Expi293-F cells
Expi293 Expression Medium
ExpiFectamine Transfection Kit
pcDNA 3.4 vector
Antibody-expressing positive control vector
Opti-MEM I
FreeStyle 293 Expression Systemlow-density, large-scale
protein expression
FreeStyle 293-F cells
FreeStyle 293 Expression Medium
293Fectin Transfection Reagents
Opti-MEM I
42
FreeStyle MAX CHO Expression systemlarge-scale protein

expression in CHO cells
FreeStyle CHO-S cells
FreeStyle CHO Expression Medium
FreeStyle MAX Transfection Reagents
OptiPro SFM

Mammalian cell lines adapted to a variety of media formulations to meet your protein expression needs.
Each lot is tested for cell growth and viability post-recovery from cryopreservation, and contains documented lineage from a low-passage Master Cell Bank. In addition, Master Seed Banks have been tested for
contamination with bacteria, yeasts, mycoplasmas, and viruses and have been characterized by isozyme
and karyotype analysis.
Source
Cell type
Adapted media
Cat. Nos.
Mammalian
293-F
FreeStyle 293
and CD 293
R79007, 11625019
Suspension*
cGMP-banked
Mammalian
293-F
CD 293
11631017
Mammalian
DG44 (Cells and

Media Kit)
CD DG44
A1100001
Mammalian
CHO-S (Cells and

Media Kit)
CD CHO
A1369601
* It is possible to achieve adherence in these cell lines by the addition of FBS or through the use of specially formulated media. Contact Technical Support for information.
FBS
Cell culture
Serum processed from bovine and other animals is a complex of over 10,000 components, including
growth factors, proteins, lipids, minerals, and attachment factors, all of them useful for growth and
maintenance of cells. Serum is a very cost-effective supplement added to basal media to make complete
media for cell culture.
The quality of serum is determined by its ability to grow healthy cells consistently over time and passages.
Factors like sterility level, absence of microorganisms, level of endotoxin, ability to grow cells without
downstream effects, and analysis of hormonal and biochemical profile of sera are important to scientists.
Gibco sera are processed within a fully integrated operations system, controlled and supervised by
Life Technologies personnel, right from the collection stage to processing to the cell culture hood. We
process sera under stringent cGMP conditions at our ISO-certified, FDA-registered facilities.
Most Gibco sera are labeled for in vitro diagnostics, compliant with the highest level of USP sterility
testing, and triple-filtered at 0.1 m.
Learn more at lifetechnologies.com/fbs.
Our Gibco sera portfolio is classified into five product categories to help meet the needs of researchers
based on cell types, specific assays, and downstream applications of cell culture.
Standard FBS
An excellent value product for basic research with robust cell lines. Includes Qualified FBS products
with endotoxin specification 50 EU/mL from USDA-approved regions, South America, Canada, and
EU-approved regions.
Performance FBS
Recommended for general cell culture with commonly used cell lines. Qualified FBS US-origin products
with low endotoxin, specification 10 EU/mL.
Performance Plus FBS
For cell culture with a broad range of cell lines, especially for sensitive cell lines. Certified FBS US-origin
products with the lowest endotoxin level, specification 5 EU/mL. These products undergo the highest
level of testing, including biochemical and hormonal profiles, useful for analysis of downstream assays.
43

Choose the FBS thats right for your project and your budget. Find this interactive table online at lifetechnologies.com/fbs.
Standard
Sera for cell culture with robust
cell lines. Good value for basic
research.
Performance
Low-endotoxin sera for general
cell culture with common cell
lines.
Performance Plus
Lowest endotoxin for a broad
range of cell types, especially
sensitive cell lines.
Product
FBS Qualified (USDA-approved,

Canada, South America origin)
FBS Qualified (US origin)
FBS Certified (US origin)
Product use
IVD
IVD
IVD
Endotoxin specification
50 EU/mL
10 EU/mL
5 EU/mL
Quality/performance testing
(gamma-irradiated upon
request)
Standard testing
Standard testing
Standard testing, plus analytical

tests for hormone and biochemical
profiles
Popular Cat. No. (500 mL)
10437028 FBS Qualified

(USDA-approved)
26140079 FBS Qualified

(US origin)
16000044 FBS Certified

(US origin)
Convenient options
Heat-inactivated
1001,000 mL sizes
50 mL One Shot packs
Heat-inactivated
1001,000 mL sizes
Heat-inactivated
1001,000 mL sizes
Secure FBS
Sera for cell culture with the least viral risk, sourced from BSE-free regions. Preferred for academic, industrial,
and preclinical research requiring low BVDV.
Cell culture
Exclusive screening for BVDV at collection

Endotoxin 10 EU/mL for FBS and donor bovine sera
Available as heat-inactivated FBS for ready-to-use convenience
Pack sizes from 100 to 1,000 mL
Recommended Gibco sera: FBS, Qualified, Australia-origin (Cat. No. 10099141), and heat-inactivated version
(Cat. No. 10100147).
Find out more about secure FBS at lifetechnologies.com/securefbs.
Specialty sera
Sera specifically qualified for cell culture for specialty research and specific assays, including stem cell
research, immunoassays, antibody production.
Embryonic stem cellqualified FBS

Charcoal-stripped FBS
Dialyzed FBS
Find out more at lifetechnologies.com/specialtysera.
Other animal sera
These products include bovine, horse, newborn calf, goat, rabbit, porcine, and chicken sera, predominantly
sourced from New Zealand, in pack sizes from 50 mL One Shot aliquots to 1,000 mL bottles.
Find out more at lifetechnologies.com/otheranimalsera.
Our animal serum products are offered in regular as well as heat-inactivated formats, in pack size options from
the convenient 50 mL One Shot aliquots to 1,000 mL, up to 20 L packs for high-throughput cell culture. In addition, several custom-processed products are available for specific research needs, including gamma-irradiated
and cell linescreened.
iMATCH lot matching tool
We also offer our free, proprietary multiparametric sera lot matching tool, the iMATCH tool, to help
you find the most consistent or highest-performing lot of serum available. Find the iMATCH tool at
lifetechnologies.com/imatch.
44
Reagents
From beginning to end, Gibco cell culture reagents are manufactured according to the highest quality and
service standards. From the most basic formulations to the newest innovations, Gibco customer-focused
innovation helps ensure our products provide superior performance and consistencyfor results that you
can count on every day.
Find more information on the reagents in this section at lifetechnologies.com/culturereagents.
Balanced salt solutions
Balanced salt solutions can provide an environment that maintains the structural and physiological integrity
of cells in vitro. All Gibco DPBS, PBS, HBSS, and EBSS balanced salt solutions are manufactured in stateof-the-art cGMP, ISO-certified facilities to offer the highest quality and consistency for reproducible results.
Tests include osmolality, pH, stability, sterility, and endotoxin tests. All powdered balanced salt solutions are
produced without sodium bicarbonate to increase stability.
HEPES buffer
HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic organic chemical buffering

agent that is widely used to provide supplemental buffering to cell culture medium at pH 7.27.6, for example,
when cell culture requires extended periods of manipulation outside of a CO2 incubator. The level of HEPES
in cell culture media generally varies from 10 mM to 25 mM. In Gibco DMEM, it is at a concentration of
25 mM; in Gibco DMEM/F-12 it is at 15 mM.
Cell culture
Sodium bicarbonate
Sodium bicarbonate (NaHCO3) is the most commonly used buffer for cell culture media. This is due to its
nutritional benefits despite the reduced buffering capacity at physiological pH.
Amino acids
Addition of amino acid supplements helps prolong the viability of cells in culture, stimulates growth, and
reduces the biosynthetic burden on cultured cells. The nonessential amino acids in this solution are prepared
in distilled water and provided at 100X concentration.
Water
We produce membrane-filtered and endotoxin-screened distilled water in a number of packaging configurations and volumes for routine cell culture. We also offer Gibco Water for Injection (WFI) for Cell Culturea
high-quality, cell culturegrade water that meets the United States Pharmacopeia monograph for water for
injection packaged in bulk for commercial use elsewhere. Gibco Water for Injection (WFI) for Cell Culture is
manufactured in ISO-certified, cGMP, and FDA-registered facilities.
Supplements
Media supplements are included in mammalian cell culture systems to help customize the growth
conditions, improve the cell viability and growth, and keep cells healthier longer. We produce an extensive selection of media supplements that help ensure the reliability and consistency of your cell culture
research.
Find out more at lifetechnologies.com/mediasupplements.
Antifoaming agent
FoamAway Irradiated AOF is a ready-to-use antifoaming agent designed to offer high performance and
convenience in combating foam in your cell culture system. It is prediluted, presterilized, and packaged to
connect directly to your bioreactor with no further processing required.
Find out more at lifetechnologies.com/foamaway.
45

Antibiotics and antimycotics
For researchers, two types of cell culture contamination require careful monitoring: the contamination of
cell cultures with microbiological organisms and the contamination of one cell line with another. Both forms
of contamination are extremely prevalent and cannot be underestimated. The decision to use antibiotics and
antimycotics to prevent contamination should be based on the individual researchers needs and experience.
Life Technologies manufactures a wide array of antibiotics and antimycotics for contamination due to
bacteria, fungi, or yeasts. While researchers may attempt to eliminate or control contaminations with antibiotics or antimycotics, it should be noted that these substances can be toxic to certain cell lines. Before using,
it is recommended that you first perform a dose-response test to determine the level at which toxicity begins.
Selection antibiotics
Gibco high-quality selection agentsprovide unique solutions for your research needs, such as dual selection and rapid stable cell line establishment. Choose the selection antibiotic that is best suited to your needs.
Cell culture
Eukaryotic selection antibiotics. Find this interactive table online at lifetechnologies.com/selectionantibiotics.

Selection antibiotic
Most common selection

usage
Common working
concentration
Available powder sizes
Available liquid sizes
Blasticidin
Eukaryotic and bacterial
120 g/mL
50 mg
10 x 1 mL, 20 mL
Geneticin (G-418)
Eukaryotic
100200 g/mL bacteria

200500 g/mL mammalian cells
1 g, 5 g, 10 g, 25 g
20 mL, 100 mL
Hygromycin B
Dual-selection experiments and eukaryotic
200500 g/mL
20 mL
Mycophenolic acid
Mammalian and bacterial
25 g/mL
500 mg
Puromycin
0.25 g/mL
10 x 1 mL, 20 mL
Zeocin
Mammalian, insect, yeast,

bacterial, and plant
50400 g/mL
8 x 1.25 mL, 50 mL
Bacterial selection antibiotics. Find this interactive table online at lifetechnologies.com/selectionantibiotics.
46
Selection antibiotic
Most common selection

usage
Common working
concentration
Available powder sizes
Available liquid sizes
Actinomycin D
Bacterial
1 g /mL
5 mg, 10 mg
Ampicillin, sodium salt
Bacterial
1025 g/mL
200 mg
Blasticidin
120 g/mL
50 mg
10 x 1 mL, 20 mL
Carbenicillin, disodium
salt
Agrobacterium and E. coli
100500 g/mL
5g
Kanamycin sulfate
Bacterial
100 g/mL
5 g, 25 g
100 mL
Mycophenolic acid
Mammalian and bacterial
25 g/mL
500 mg
Neomycin sulfate
Bacterial
50 g/mL
100 g
Polymyxin B sulfate
Bacterial
100 units/mL
25 MU
Puromycin
0.25 g/mL
10 x 1 mL, 20 mL
Streptomycin sulfate
Bacterial
50100 g/mL
100 g
Zeocin
Mammalian, insect, yeast,

bacterial, and plant
75400 g/mL
8 x 1.25 mL, 50 mL
Growth factors
Gibco growth factors, chemokines, and cytokines are pure, high-quality proteins bioassayed with Gibco media, making them
compatible with the cell culture media brand you use and trust. In addition, each protein is analyzed for purity along with structural
homogeneity to help ensure a biologically active protein is produced.
Find more information online
On our website, youll find details on our broad selection of growth factors, cytokines and chemokines, including VEGF, EGF, FGFb,
NGF, and SCF. Go to lifetechnologies.com/growthfactors to find:
Product groups narrowed alphabetically by name or by protein family or species

Data demonstrating the purity, biological activity, and functionality of our recombinant proteins
Information on our ISO 13485quality growth factors for clinical research
Insect cell culture

Insect cell culture is a common choice for heterologous protein expression. For large-scale production or
basic research, insect cells are able to express large quantities of protein with complex posttranslational
modifications. Gibco insect media have been formulated for maximum growth and protein yields. These
media, in combination with Gibco preadapted cell lines, will provide a convenient system to save you time
and effort.
Cell culture
Insect cell media selection guide. Find this interactive table online at lifetechnologies.com/insectcellculture.
Basic insect
cell research
Serum-free growth and protein

expression
Highest consistency and

protein expression
Graces Insect Medium
Sf-900 II
Sf-900 III
Serum free
Protein free
Source origin
Animal
Serum-free
Animal originfree
Low
(FBS variability)
Medium
(medium hydrolysate concentration)
High
(low hydrolysate concentration)
Optimized to preventfoaming
Compatible insectcells
Sf9 and Sf21
Sf9 and Sf21
Sf9 and Sf21
Supported cell density

(Sf9cells/mL)
3 x 106 cells/mL
10 x 106 cells/mL
1014 x 106 cells/mL
Packaging
100 mL, 500 mL, 10 x 1L (powder)
500 mL, 1,000 mL, 5 L (bags),

10 L (bags), 20 L (bags)
500 mL, 1000 mL, 10 L (bags),

20 L (bags)
Drosophila S2 Cell Culture
Drosophila S2 cells are used for heterologous protein expression

in the Drosophila Expression System (DES). The S2 cell line was
derived from a primary culture of late stage Drosophila melanogaster embryos (Schneider, 1972). This adaptable cell line grows
without CO2, at room temperature, as a semi-adherent monolayer or suspension in Schneiders Drosophila Medium. Learn
more at lifetechnologies.com/insectcellculture.
High Five Cell Culture
High Five Cells (BTI-TN-5B1-4) are a clonal isolate, derived

from the parental Trichoplusia ni cell line. They are optimized for
the growth and maintenance of cells used with the Baculovirus
expression vector system (BEVS) for adherent or suspension
culture. Adapted to serum-free suspension culture in Gibco
Express Five serum-free medium.
Learn more about Express Five SFM (Cat. No. 10486025).
47

Insect cell lines adapted to a variety of media formulations to meet your protein expression needs. Each lot is tested for cell
growth and viability post-recovery from cryopreservation, and contains documented lineage from a low-passage Master Cell
Bank. In addition, Master Seed Banks have been tested for contamination with bacteria, yeasts, mycoplasmas, and viruses
and have been characterized by isozyme and karyotype analysis.
Source
Cell type
Adapted media
Cat. Nos.
Suspension
Adherent
Insect
Sf9
Graces, Sf-900 II, and

Sf-900 III
B82501, 11496015,
12659017
Insect
Sf21
Graces, Sf-900 II, and

Sf-900 III
B82101, 11497013,
12682019
Insect
High Five
Express Five SFM
B85502
Drosophila S2
Schneiders
R69007
Insect
(Semi-adherent)
Neurobiology media
We offer an array of media for neural cell culture, including neural primary and neural stem cells, media,
supplements, substrates, and growth factors. All of our products have been developed to work together for
optimal performance as a complete cell culture system. Gibco media and supplements are the most widely
used for neural cell culture. Gibco products are designed to provide the highest quality, consistency, and
performancefor results you can count on.
Cell culture
Neurobasal media
Neurobasal media are a full line of basal media formulated to sustain optimal culture of neurons. Our
Neurobasal media must be supplemented with either a serum-free supplement (such as B-27 Supplement
or N-2 Supplement) or serum, and 0.5 mM L-glutamine or GlutaMAX-I Supplement.
Cited in many peer-reviewed publications

Supports growth of neurons from the hippocampus, embryonic striatum, substantia nigra, septum,
cortex, neonatal dentate gyrus, and cerebellum when combined with B-27 Supplement
Choose the right Neurobasal media for your experimental needs from the list below:
Requirements
Recommended products
For prenatal and fetal neurons
Neurobasal Medium (1X) Minus Phenol Red (Cat. No. 12348017)

Neurobasal Medium (1X) (Cat. No. 21103049)
For postnatal and adult brain neurons
Neurobasal-A Medium (1X) Minus Phenol Red (Cat. No. 12349015)

Neurobasal-A Medium (1X) (Cat. No. 10888022)
Neurobasal-A Medium (1X) (Custom) (Cat. No. 0050128DJ)
For tumor cell lines

Neurobasal-A Medium (1X) (Cat. No. 10888022)
No glucose/no sodium pyruvate
No phenol red
Neurobasal Medium (1X) Minus Phenol Red (Cat. No. 12348017)

Learn more at lifetechnologies.com/neurobasal.
48

Hibernate media
Hibernate media, when supplemented with B-27 Supplement and GlutaMAX-I Supplement, allow for
the manipulation of neurons at ambient CO2 for at least 48 hours while retaining their viability. Hibernate
media can also be used to preserve viable brain tissue for up to a month when stored at 4C and have
also been shown to be suitable transport media to ship various tissues and biological specimens, including
umbilical cord tissue. There are two types of Hibernate media, Hibernate-A and Hibernate-E, formulated
to be used for postnatal and embryonic neurons, respectively. These two products have similar formulations
except for a difference in osmolality: Hibernate-A has a higher range of osmolality than Hibernate-E.
Learn more at lifetechnologies.com/hibernate.
Astrocyte medium
Gibco Astrocyte Medium (Cat. No. A1261301) has been specially formulated to support the growth of
primary human and rat astrocytes while retaining their phenotypes. The medium has been internally validated and optimized to provide an optimal balance between cell growth and the maintenance of cell viability,
normal phenotype, and function. If more rapid proliferation is desired, addition of an appropriate amount of
epidermal growth factor (EGF) is recommended, although cells may exhibit morphological and phenotypic
changes.
Extracellular matrices
Cell culture
Culturing cells on flat plastic surfaces results in artificial two-dimensional sheets of cells. In the in vivo
state, human cells experience a three-dimensional environment, completely surrounded by other cells,
membranes, fibrous layers, and adhesion proteins. Our Gibco extracellular matrices, scaffolds, and
proteins produce a growth environment closer to that found in vivo, and this physiologically relevant environment allows you to get more realistic cell biology data.
At lifetechnologies.com/3dcellculture, you can find information about:
Rat tail and bovine collagen I

Natural mouse laminin to facilitate the attachment and expansion of ESCs, iPSCs, and NSCs
Extracellular matrix (ECM) proteins, including fibronectin and vitronectin
Geltrex LDEV-free matrix products
Geltrex Matrix is a soluble form of reduced growth factor (RGF) basement membrane extract (BME) purified
from murine Engelbreth-Holm-Swarm tumor.
Free of the lactate dehydrogenase-elevating virus (LDEV), making it ideal for all types of cell culture and
mouse in vivo research
Tested for its ability to support endothelial tube formation from cryopreserved endothelial cells
Available in standard and hESC-qualified formulations
Learn more at lifetechnologies.com/geltrex.
AlgiMatrix 3D cell culture system
The AlgiMatrix 3D Culture System is the first user-friendly, animal-free bioscaffold available for the development of higher-fidelity cell culture models that are more predictive of disease states and drug responses.
Three-dimensional, porous alginate cell culture platform

Chemically defined, animal originfree material
Supports formation of 3D cellular aggregates that more closely reflect normal cell morphology and
behavior
Easy visualization of cells

Lyophilized product available in 6-well, 24-well, and 96-well plate formats
Learn more at lifetechnologies.com/algimatrix.
49
Microbial culture
Microbial culture is a well-established research tool in molecular biology for the culture of bacteria and
yeasts. Life Technologies offers a wide array of bacterial culture and yeast culture products, including growth
media and reagents, to help you achieve your desired results.
Learn more at lifetechnologies.com/microbialculture.
Bacterial
Liquid and powder growth media specifically formulated to culture bacteria for use in applications including cloning
and protein expression.
Choose your type of bacterial growth medium.

LB broth and LB agar
Most referenced bacterial medium for growth and maintenance of recombinant E. coli strains
Terrific broth
Nutritionally rich bacterial medium for higher-density growth and maintenance of recombinant E. coli strains
M9 minimal
Commonly used bacterial medium for cultivation and maintenance of E. coli strains
MagicMedia Medium
Customized bacterial medium for growth of T7-regulated E. coli strains for autoinduction of protein expression
ImMedia Medium
Premixed, presterilized bacterial medium containing selection antibiotics for growth of E. coli strains
Other media
Microbial media for outgrowth of transformed E. coli cells (S.O.C. medium), solid microbial media (Select
Agar), and growth of hosts for replication of M13 vectors (2-YT Broth)
Cell culture
Learn more at lifetechnologies.com/bacterialmedia.
Yeast
Liquid and powder growth media specifically formulated to culture yeast for use in maintaining and propagating yeast strains.
Choose the appropriate yeast growth medium.

PichiaPink Media
Prepackaged media pouches for use with PichiaPink Yeast Expression System, containing media to grow
P. pastoris yeast strains for transformation and selection
CSM medium
Specialty medium for making agar plates to grow S. cerevisiae (MaV203) competent cells
YPD broth
Commonly used yeast medium for maintenance and propagation of P. pastoris and S. cerevisae yeast strains
Yeast Nitrogen Base (YNB)
Base medium for preparation of minimal and synthetic defined yeast media
Learn more at lifetechnologies.com/yeastmedia.
Algae
Gibco TAP Growth Medium (Cat. No. A1379801) optimized for Chlamydomonas reinhardtii, and Gibco BG-11
Growth Medium (Cat. No. A1379901) optimized for cyanobacteria.
Learn more about algae protein expression systems at lifetechnologies.com/algaekit.
50
Cytogenetics
Gibco cytogenetics products are specifically formulated to help deliver clear, reproducible results and are application-tested by an independent laboratory. Each medium
has been designed and optimized for the analysis of amniotic fluid cells, chorionic
villus samples, bone marrow cells, or peripheral blood lymphocytes.
Optimized and prequalified for cytogenetics

Provide high mitotic index
Deliver excellent chromosomal morphology
Offer clear, reproducible results that are easy to analyze and interpret
Product use: IVD
Learn more at lifetechnologies.com/cytogenetics.
AmnioMAX media
Gibco AmnioMAX-C100 and AmnioMAX II complete media are designed to

increase cell attachment and growth rates and provide high metaphase yields.
Learn more at lifetechnologies.com/amniomax.
MarrowMAX Bone Marrow Medium
Cell culture
A bone marrow aspirate provides unique and valuable research data, but cell numbers
are generally very lowtherefore, the medium you select could not be any more critical. MarrowMAX Bone Marrow Medium contains a novel stromal cellconditioned
medium for optimal growth and minimizes the need to supplement media, helping
to save both time and money. MarrowMAX medium far outperforms commercially
available formulations supplemented with giant cell tumor (GCT)conditioned media,
offering consistent lot-to-lot performance with a higher mitotic index and superior
chromosome morphology.
Learn more at lifetechnologies.com/marrowmax.
PB-MAX Karyotyping Medium
PB-MAX Karyotyping Medium is a fully supplemented, RPMI 1640based medium

optimized for the karyotype analysis of peripheral blood lymphocytes. Ready to use, it
is proven for performance and reliability.
Learn more at lifetechnologies.com/pbmax.
KaryoMAX products
The KaryoMAX line of products is designed to reduce culture time and days to
harvest. Our line of KaryoMAX reagents includes KaryoMAX Colcemid, KaryoMAX
Giemsa Stain, and KaryoMAX potassium chloride solution.
Learn more at lifetechnologies.com/karyomax.
51
Passage
Gibco cell dissociation products are ideal for use with tissues and cell monolayers. Gibco trypsin and TrypLE
cell dissociation reagents come in a wide variety of formats to help meet the diverse needs of researchers
performing adherent cell culture.
Cell dissociation reagent selection guide. Find this interactive table online at lifetechnologies.com/celldissociation.
Fast, general
purpose
Gentle and convenient

for research use
Gentle and convenient

for bioproduction/
industrial applications
Gentle and fast for

strongly adherent
cells
Specialized for neural

and pluripotent stem
cell dissociation
Trypsin
TrypLE Express (1X)
TrypLE Select (1X)
TrypLE Select (10X)
StemPro Accutase
Source origin
Animal origin
(porcine)
Animal originfree
Animal originfree
(dedicated AOF
machinery)
Animal originfree
(dedicated AOF
machinery)
Serum-free, animal
origin (marine invertebrate)
Storage temperature
Frozen
(5 to 20C)
Ready to use (room

temperature stable)
Ready to use (room

temperature stable)
Ready to use (room

temperature stable)
Refrigerated
(2 to 8C)
Higher cell viability
Requires trypsin
inhibitors/neutralizers
Inhibition by dilution
(no inhibitors needed)
Available packaging
100 mL, 500 mL,

100 g powder
100 mL, 500 mL,

5 L bags
100 mL, 500 mL
100 mL, 500 mL
100 mL
Cell culture
Inactivation method
52
Automated cell counting

The Countess Automated Cell Counter takes the subjectivity and tedium out of one of the fundamental steps of cell culture
counting cells and assessing viability. The Countess Automated Cell Counter helps to eliminate subjectivity and minimize human
error for comprehensive, reproducible data.
The Countess Automated Cell Counter is:
Accurateeliminate the subjectivity of manual cell counting; no guessing, no user-to-user variability

Fastcounts live and dead cells and measures viability and average cell size, typically in 30 seconds with just 10 L of sample
Convenientno setup, cleaning, or service required
Easy workflow and accurate results
The Countess Automated Cell Counter is easy to use. Simply pipet the sample into the counting slide, insert the slide into the
Countess Automated Cell Counter, then press Count cells; results are displayed in 30 seconds.
See the Countess Automated Cell Counter in action at lifetechnologies.com/countess.
Cell culture
The Countess Automated Cell Counter workflow. (A) Pipet the sample into the counting slide. (B) Insert the slide into the Countess Automated Cell Counter. (C) Press
Count cells. (D) Results are displayed in 30 seconds.
53
Human primary cells

Our portfolio of Gibco primary cell cultures and complete primary cell culture systems (including cells, media and supplements,
matrices, cryopreservation medium, and analysis tools) give you the potential to generate more physiologically relevant data.
Gibco complete cell culture systems are:
Optimized for each cell type

Easy to use, offering single-shot supplementation
Available in custom solutions for unique research needs
We maintain stocks of the following primary cells:
Keratinocytes
Fibroblasts
Melanocytes
Corneal epithelial cells
Microvascular entothelial cells
Smooth muscle cells
Large vessel endothelial cells (aortic, pulmonary, and umbilical)
Hepatocytes
Astrocytes
Skeletal myoblasts
Mammary epithelial cells
Primary cells
Browse our entire selection human primary cells at lifetechnologies.com/primarycells.
Custom primary cells and media
Life Technologies welcomes requests for custom preparations of cell culture products and contract research. Please contact
your account manager or technical sales specialist, and we will work with you to develop a solution that meets your research and
budgetary needs.
Custom cell culture products and services
Custom cell isolations and configurations

Custom medium and supplement formulations
Cell pellets suitable for RNA isolation and other purposes
Additional cell characterization and virus testing
Research applications for primary cells

Basic structure/function
studies
Cancer biology
Drug discovery/cosmetics/beauty
and personal care studies
In vitro alternatives to
animal testing
Regenerative medicine/
therapy
Dermal modeling
Angiogenesis
Acne
Corrosivity
Burn therapy
Gene regulation
Melanoma
HTS/HCA screening
Cosmetics and topicals
Chronic skin ulcers
Signal transduction
Normal controls
Pigmentation
Household product irritancy
Cosmetic (wrinkles, scars,

hair growth)
Secondary and tertiary screenings
Safety assessment testing

services and products
Wound healing
Cell co-culturing
Toxicology screening
54

Products for engineering and analyzing primary cells
Life Technologies offers a host of products that enable further engineering of primary cells, as well as analysis of activity and
function within primary cell models. These include detection technologies such as CellLight and Premo reagents based on
BacMam technology, which enable efficiently delivery and gene expression in mammalian cells, even for the most difficult-totransfect primary cells. Additionally, our instrument platforms can simplify everything from counting, transfecting, and visualizing
primary cells. These platforms include:
Neon Transfection System (lifetechnologies.com/neon)

Tali Image-Based Cytometer (lifetechnologies.com/tali)
FLoid Cell Imaging Station (lifetechnologies.com/floid)
Attune Acoustic Focusing Cytometer (lifetechnologies.com/attune)
Cultured in EpiLife Medium + HKGS
Cultured in Medium 154 + HKGS
Primary cells
HEKn, day 1
HEKn, day 3
HEKn, day 5
Comparison of HEKn (Cat. No. C-001-5C), secondary culture, grown in either EpiLife Medium or Medium 154.
HCEC Growth Rates
B
Population doublings/day
1.2
3 culture
5 culture
1.0
7 culture
0.5
KSFM
dKSFM + CM
Growth Medium
Fluorescent multiplex imaging of microfilaments in human dermal fibroblasts.

(A) Phalloidin Alexa Fluor 488 (green); Hoechst 33342 (blue). (B) Antialpha/
tubulin/goat anti-rabbit Alexa Fluor 555 (red); phalloidin Alexa Fluor 488 (green);
Hoechst 33342 (blue).
HCEC were thawed and seeded according to product

instructions in Keratinocyte SFM (KSFM) or Defined
Keratinocyte SFM (dKSFM). Cells were passaged at
90% confluence, and population doublings per culture
were calculated. Bars show the mean growth in triplicate T-25 flasks, with standard deviation.
55

Life Technologies human primary cell systems quick reference guide
Anatomical location of
derivation
Cells
Acronym
Cryopreserved
product Cat. No.
Skin
Human Epidermal Keratinocyte, neonatal (animal productfree)
HEKn-APF
C0205C
Human Epidermal Keratinocyte, adult (animal productfree)
HEKa-APF
C0215C
Human Epidermal Keratinocyte, neonatal
HEKn
C0015C
Human Epidermal Keratinocyte, adult
HEKa
C0055C
Human Epidermal Keratinocytes, pooled
HEKp
A13401
Human Melanocyte-neonatal, lightly pigmented
HEMn-LP
C0025C
Human Melanocyte-neonatal, moderately pigmented
HEMn-MP
C1025C
Human Melanocyte-neonatal, darkly pigmented
HEMn-DP
C2025C
Human Melanocyte-adult, lightly pigmented
HEMa-LP
C0245C
Human Dermal Fibroblasts, neonatal
HDFn
C0045C
Human Dermal Fibroblasts, adult
HDFa
C0135C
Human Microvascular Endothelial Cell, neonatal dermis
HMVECnd
C0105C
Human Microvascular Endothelial Cell, adult dermis
HMVECad
C0115C
Human Aortic Endothelial Cell
HAEC
C0065C
Human Pulmonary Artery Endothelial Cell
HPAEC
C0085C
Human Aortic Smooth Muscle Cell
HASMC
C0075C
Human Coronary Artery Smooth Muscle Cell
HCASMC
C0175C
Primary cells
Heart
Human Pulmonary Artery Smooth Muscle Cell
HPASMC
C0095C
Cornea
Human Corneal Epithelial Cell
HCEC
C0185C
Quadriceps
Human Skeletal Myoblasts, small size
HSkM
A12555
Human Skeletal Myoblasts, large size

Umbilical cord
A11440
Human Umbilical Vein Endothelial Cell
HUVEC
C0035C
Human Umbilical Vein Endothelial Cell (pooled, 500,000 cells)
HUVEC
C0155C
Human Umbilical Vein Endothelial Cell (pooled, 1 million cells)
HUVEC
C01510C
Breast
Human Mammary Epithelial Cell
HMEC
A10565
Brain
Gibco Human Astrocytes
NA
N7805100
Gibco Human Astrocyte Kit
N7805200
* Also available in Calcium Free and Calcium Free/Phenol Red Free Kits: EpiLife CF (Cat. No. MEPICF500) and CF/PRF (Cat. No. MEPICFPRF500), Medium 154 CF (Cat. No. M154CF500) and
CF/PRF Kits (Cat. No. M154CFPRF500)
Requires plating with Coating Matrix (Cat. No. R011K) for efficient cell attachment
Also available as Calcium Free Kit: Medium 254CF Kit (Cat. No. M254CF500)
Animal productfree supplements
** Also available as a Phenol Red Free: Medium 200PRF (Cat. No. M200PRF500)
Requires plating with Geltrex matrix for efficient cell attachment.
56
Growth medium
Cat. No.
Growth supplement
Acronym
Cat. No.
EpiLife*
MEPI500CA
S7
S7
S0175
EpiLife*
MEPI500CA
EpiLife Defined Growth Supplement
EDGS
S0125
Human Keratinocyte Growth Supplement
HKGS
S0015
Human Keratinocyte Growth Supplement Kit
HKGS kit
S001K
Medium 154*
M154500
Human Keratinocyte Growth Supplement
HKGS
S0015
Human Keratinocyte Growth Supplement Kit
HKGS kit
S001K
Medium 254
M254500
Human Melanocyte Growth Supplement
HMGS
S0025
Medium 254
M254500
Human Melanocyte Growth Supplement-2
HMGS-2
S0165
Medium 106
M106500
Low Serum Growth Supplement
LSGS
S00310
Low Serum Growth Supplement Kit
LSGS Kit
S003K
M131500
Microvascular Growth Supplement
MVGS
S00525
Medium 200**
M200500
Large Vessel Endothelial Supplement
LVES
A1460801
Medium 231
M231500
Smooth Muscle Cell Growth Supplement
SMGS
S00725
Smooth Muscle Cell Differentiation Supplement
SMDS
S0085
KSFM
17005042
dKSFM
10744019
DMEM, Low Glucose, Pyruvate
11885084
2% Horse Serum
Medium 200**
M200500
Large Vessel Endothelial Supplement
LVES
A1460801
Medium 171
M171500
Mammary Epitehlial Growth Supplement
MEGS
S0155
HuMEC Ready
12752010
Note: Supplements included with media
HuMEC Basal Serum Free Medium
12753018
HuMEC Supplement Kit
NA
12755013
Gibco Astroctye Medium
A1261301
Primary cells
Medium 131 (w/ Attachment Factor)
Note: Supplements included with media

16050130
NA
NOTE: Included in kit
57

Life Technologies human primary cell systems quick reference guide
Anatomical location of
derivation
Cryopreserved
product Cat. No.
HEP10, Pooled Human Cryopreserved Hepatocytes

Human Suspension Hepatocytes, Metabolism Qualified, Male
Human Suspension Hepatocytes, Metabolism Qualified, Female
Human Suspension Hepatocytes, Metabolism Qualified, Male, 912 million
Human Suspension Hepatocytes, Metabolism Qualified, Female, 912 million
Human Suspension Hepatocytes, Polymorphic Donors
Human Suspension Hepatocytes, Polymorphic Donors, 912 million
Human Suspension Hepatocytes, Transporter Qualified, 48 million
Human Suspension Hepatocytes, Transporter Qualified, 912 million
Human Plateable Hepatocytes, Induction Qualified
Human Plateable Hepatocytes, Metabolism Qualified
Human Plateable Heptocytes, Transporter Qualified
Human Plateable Heptocytes, Uptake Qualified
HMCS10
HMCS1S
HMCS2S
HMCS1L
HMCS2L
HMCSPS
HMCSPL
HMCSTS
HMCSTL
HMCPIS
HMCPMS
HMCPTS
HMCPUS
Primary cells
Liver
Cells
58
Thawing reagents
Cat. No.
Plating reagents
Cat. No.
Maintenance reagents
Cat. No.
Cryopreserved
Hepatocyte Recovery
Medium (CHRM)
CM7000
Williams Medium E (1X, no phenol red)
A1217601
Williams Medium E (1X, no phenol red)
A1217601
Hepatocyte Plating Supplement Pack

(serum-containing)
CM3000
Hepatocyte Maintenance Supplement

Pack (serum-free)
CM4000
Collagen I, Coated Plates (6-well) or

Collagen I, Coated Plates (96-well) +
Geltrex LDEV-Free Reduced Growth
Factor Basement Membrane Matrix
A1142801
A1142802
A1142803
A1413202

Collagen I, Coated Plates (96-well) +
Geltrex LDEV-Free Reduced Growth
Factor Basement Membrane Matrix
A1142801
A1142802
A1142803
A1413202
Primary cells
59
Pluripotent stem cells (PSCs)

Culture
Life Technologies offers an extensive range of Gibco media, supplements, and reagents to help meet all of your stem cell culture
needs. Culture and expand stem cells in cGMP-manufactured serum-free, feeder-free, xeno-free cell culture systems. Manufactured
with superior quality standards, our products offer the consistency and accuracy you need to get the most from your experiments.
Find out about all of the options for PSC culture at lifetechnologies.com/culturepsc.
Primary and stem cells
PSC culture medium selection guide.

The standard for serum-free,
feeder-based culture of ESCs and
iPSCs from multiple species
Feeder-free culture with

flexibility to optimize
bFGF concentration and
maintain differentiation
capacity of PSCs
Optimal feeder-free
culture of all human
ESC and iPSC lines
with minimal lot-tolot variability
Fully documented xeno-free

PSC culture reducing the
burden in qualifying reagents,
facilitating transition from
bench to clinic
Product
KnockOut Serum Replacement
StemPro hESC SFM
Essential 8 Medium KnockOut CTS XenoFree

ESC/iPSC Kit
Cat. No.
10828028, 10828010
A1000701
A14666SA
A1448801, A1448701
Product use
Class II Medical Device
RUO
RUO
RUO
Recommended
applications
Multiple species (including human

and mouse); feeder culture; reprogramming on feeders; differentiation of PSCs
Human; feeder-free
culture
Human; feeder-free,
xeno-free culture;
feeder-free reprogramming
Human; feeder-based and

feeder-free xeno-free culture
for cell therapy research
Lot-to-lot variability
Medium
Medium
Low
Medium
Cell morphology
Compact colonies with smooth

edges; high nuclear-to-cytoplasmic
ratio
Compact colonies with

smooth edges; some
colonies will have undefined borders with spiky
edges
Compact colonies
with smooth edges;
high nuclear-tocytoplasmic ratio
Compact colonies with smooth

edges; some colonies will have
undefined borders with spiky
edges
Components/unit
1 x 100 mL or 500 mL
1 x 10 mL StemPro
Supplement;
1 x 500 mL DMEM/F-12
with GlutaMax-I;
1 x 40 mL 25% BSA
1 x 10 mL Essential
8 Supplement;
1 x 500 mL
DMEM/F-12 with
GlutaMax-I
1 x 100 mL KnockOut SR
XenoFree CTS
1 x 500 mL KnockOut DMEM
CTS
1 x 5 mL KnockOut SR GF
Cocktail CTS
Additional components required for

complete medium
DMEM/F-12 (1X) with

GlutaMAX-I (Cat. No. 10565018)
NEAA (Cat. No. 11140050)
bFGF (Cat. No. PHG0264)
2-mercaptoethanol
(Cat. No. 21985023)
bFGF
(Cat. No. PHG0264)
2-mercaptoethanol
(Cat. No. 21985023)
No additional compo- bFGF CTS

(Cat. No. CTP0263)
nents required
2-mercaptoethanol
(Cat. No. 21985023)
Stability of complete Stable at 4C for 2 weeks

medium
Stable at 4C for 2 weeks
Stable at 4C for 2
weeks
Specifically
designed for cell
therapy research
No
No
No
Yes
Available support
resources
Protocols for culturing PSCs, publications, hands-on training, product

finder
Protocols for culturing

PSCs
Protocols for
Protocols for culturing PSCs,
culturing PSCs, FAQs, FAQs, product documentation
hands-on training
For cost-effective option, choose the

KnockOut ESC/iPSC Media Kit
(Cat. No. A1412901)
60

KnockOut Serum Replacement (KSR) Cat. No. 10828010, 10828028
Well-referenced, proven standard
KnockOut Serum Replacement and basal media deliver confidence, with over 2,400
peer-reviewed publications.
StemPro hESC SFM Cat. No. A1000701
Flexible feeder-free culture
StemPro hESC SFM optimizes bFGF concentration and maintains the differentiation
capability of PSCs.
Essential 8 Medium Cat. No. A14666SA

Feeder-free culture for all
Essential 8 Medium contains only the necessary components for optimal PSC culture
for every step of your research.
KnockOut CTS XenoFree ESC/iPSC Kit Cat. No. A1448801, A1448701

Traceability for cell therapy research
The KnockOut CTS XenoFree ESC/iPSC Kit helps reduce the burden in qualifying
reagents, facilitating transition from bench to clinic.
KnockOut ESC/iPSC Media Kit Cat. No. A1412901, A1413001

Bundled reagents for convenience
A cost-effective and convenient way to purchase the reagents required for feeder-based
culture of human and nonhuman embryonic stem cells (ESCs) or induced pluripotent
stem cells (iPSCs). This kit includes DMEM/F-12 with GlutaMAX-I, KnockOut Serum
Replacement, and FGF-basic.
KnockOut basal media

KnockOut DMEM Cat. No. 10829018
Optimized for growth of undifferentiated ESC and iPSCs
KnockOut DMEM is a chemically defined, reduced-osmolality basal medium designed
for use with KnockOut Serum Replacement.
KnockOut DMEM/F-12 Cat. No. 12660012

Accelerates growth of ESCs and iPSCs
KnockOut DMEM/F-12 is a chemically defined, low-osmolality basal medium for use
with KnockOut Serum Replacement.
Basic Fibroblast Growth Factor (bFGF) Cat. No. PHG0264
Gibco growth factors are designed to provide the activity, stability, and validation
required for your stem cell research.
High biological activity and high puritymore results with less protein
Proven compatibilityGibco proteins are bioassayed with Gibco media
Find bFGF and other growth factors at lifetechnologies.com/growthfactors
61
Matrix products and attachment factors

We provide extracellular matrices (ECMs) that are designed to minimize adaptation time, maximize cell performance, and help
meet regulatory requirements. Whether you are culturing your cells on feeder layers or require feeder-free xeno-free conditions
for more clinical applications, Gibco products provide you with all the choices to culture your pluripotent stem cells (PSC) in the
right environment. Use the guide below to help you choose the right product for your cells.
ECM and attachment factor selection guide.
Most commonly used basement

membrane matrix for ESC and iPSC
cultures
Optimized for use with Essential 8

Medium
Optimal for clinical research applications that require human origin

only materials
Product
Geltrex LDEV free hESC Qualified
Vitronectin (VTN-N)
CELLstart CTS
Cat. No.
A1413301
A14701SA
A1014201
Product
RUO
RUO
RUO
Recommended
applications
Feeder and feeder-free culture
Feeder-free and xeno-free culture
Feeder-free, xeno-free culture
Recommended medium
pairing
StemPro hESC SFM
Essential 8 Medium
KnockOut SR XenoFree CTS

supplemented medium
Source origin
Murine
Human (recombinant)
Blend of human native proteins
Defined
No
Yes
Yes
Storage temperature
Frozen (20 to 80 C)
Frozen (20 to 80 C)
Refrigerate (2 to 8C)
Plate preparation time
1 hr incubation at 37C
1 hr incubation at room temp
12 hr incubation at 37C
Shelf life of prepared

plate
2 weeks at 28C
1 week at 28C
17 days at 28C (tightly sealed)
Unit size
1 mL or 5 mL
1 mL
2 mL
Number of plates
coated/unit
16 x 6 well plates from 1 mL vial

diluted 1:100
16 x 6-well plates from 1 mL
22 x 6-well plates, when diluted 1:50
Note: Attachment Factor (0.1% Gelatin, Cat. No. S006100) may be used for feeder-layer attachment.
62
Passaging PSC cultures

We offer options for both enzymatic and mechanical passaging of PSC cultures. Which method you choose depends on the condition of the culture at the time you want to passage and what will be done with the cells after passaging. Use the guide below to
help you select the right method.
Passaging method selection guide.

Standard
reagent
for clump
passaging in
feeder and
feeder-free
cultures
Quick and gentle

dissociation, recommended for singlecell passaging
Recommended
for clump
passaging with
feeder-free
Essential 8
Medium
Optimal
for clinical
research
applications
that require
human origin
only materials
Quick and
easy manual
passaging tool
for feeder and
feeder-free
cultures
Product
Dispase
Collagenase IV
StemPro Accutase
UltraPure 0.5 M
EDTA
TrypLE
Select CTS
StemPro
EZPassage
Disposable Stem
Cell Passaging
Tool
Cat. No.
17105041
17104019
A1110501
15575020
A1285901
23181010
Recommended
applications
Passaging in
clumps
Passaging in
clumps
Dissociation to single
cells
Passaging in
clumps
Single-cell
passaging
Mechanical
passaging in
clumps
Source origin
Bacillus polymyxa
Clostridium
histolyticum
Marine invertebrate
Chemical, AOF
Recombinant
enzyme (AOF)
NA
Form
Lyophilized
powder
Lyophilized
powder
Ready-to-use liquid
Liquid, dilute
1,000x
Ready-to-use
liquid
Individually
wrapped
Storage temperature
28C
28C
28C
Room
temperature
Room
temperature
Room
temperature
Shelf life of liquid

format
2 weeks
2 weeks
2 years
2 years
2 years
NA
Inactivation method
Inhibition by
dilution
(no inhibitors
needed)
Inhibition by
dilution
(no inhibitors
needed)
Inhibition by
dilution
(no inhibitors
needed)
Inhibition by
dilution
(no inhibitors
needed)
NA
Unit size
5g
1g
100 mL
4 x 100 mL
100 mL
Box of 10 units
Optimized
concentration for use
2 mg/mL
1mg/mL for
feeder-dependent, 10 mg/mL
for feeder-free
1X solution
0.5 mM
1X solution
NA
Number of plates
passaged/unit
1,250 x 60 mm
plates
500 x 60 mm
plates at
1 mg/mL,
50 x 60 mm
plates at
10 mg/mL
50 x 60 mm plates
50,000 x 60 mm
plates
(100 mL at 1,000X
dilution)
50 x 60 mm
plates
10 plates
Recommended
for clump
passaging in
feeder and
feeder-free
cultures
63
Tools for reprogramming

Selecting the right reprogramming tool to generate induced pluripotent stem cells (iPSCs) can leave you
with questions. And no one has more answers than Life Technologies. From traditional methods (lentivirus) to nonintegrating technologies (Sendai virus and episomal vectors), Invitrogen products provide the
breadth and depth you depend on. That means more solutions to help meet your safety, efficiency, and
budgetary needs.
Learn more at lifetechnologies.com/cellreprogram.
Lentiviral iPSC reprogramming particles

Well-referenced, flexible technology
Choose from eight different high-titer lentiviral iPSC reprogramming particles

for your custom iPSC experiments.
Learn more at lifetechnologies.com/lenti.
Episomal reprogramming vectors Cat. No. A14703

Safety with confidence
Create reproducible iPSCs from normal and diseased cell types with Invitrogen
episomal vectors.
CytoTune-iPS Sendai Reprogramming Kit

High efficiency for difficult cell types
Reprogram challenging cell types with the easy-to-use, nonintegrating CytoTune-iPS

Sendai Reprogramming Kit.
Learn more at lifetechnologies.com/cytotune.
Reprogramming tool selection guide.
64
Well-referenced technology for iPSC

generation from fibroblasts
Proven technology for viral-free

iPSC generation from normal and
diseased cell types
Efficient generation of
integration-free iPSCs from
multiple somatic cell types
Product
Lentiviral iPSC reprogramming

particles
Episomal iPSC Reprogramming

Vectors
CytoTune-iPS Sendai
Reprogramming Kit
Cat. No.
A1357101, A1357201, A1357301,

A1357401, A1357501, A1357601,
A1357701, A1357801
A14703
A1378001, A1378002
Reprogramming efficiency
0.0010.01%
0.0020.08%
0.011%
Genomic integrationfree
No
Yes
Yes
Virus-free reprogramming
No
Yes
No
Hands-on time
One application, selection and QC of

putative colonies required
One application, minimal QC of

putative colonies required
One application, minimal QC

of putative colonies required
Blood cell reprogramming
No
No
Yes
Convenience
hOct4-GFP and CMV-GFP reporters

available for transduction and
expression verification
Premixed, 3-plasmid kit; requires

Neon Transfection System (or
similar device)
Kit contains all 4 genes

required; cannot be
purchased separately
Available support resources
Standard protocol for fibroblast

reprogramming
Protocols for fibroblast

reprogramming
Protocols for fibroblast,

CD34+, PBMC
reprogramming.
FAQs, webinar, publications
Other reprogramming tools

TaqMan iPSC Sendai Detection Kit Cat. No. A13640
These TaqMan probes are used for detecting the presence and determining the levels
of Sendai virus and exogenous transcription factors (OCT 3/4, SOX2, KLF4, and c-Myc)
delivered by the Sendai virus from the CytoTune-iPS Sendai Reprogramming Kit. The
primers in the TaqMan iPSC Sendai Detection Kit will not detect the corresponding
endogenous factors.
Primary cells for reprogramming

Primary cells are cells taken directly from living tissue and established for growth
in vitro. These cells have undergone very few population doublings and are therefore
more representative of the main functional component of the tissue from which they
are derived
Find all of our primary cells at lifetechnologies.com/primarycells.
Fibroblasts
Primary human dermal fibroblasts from either adult or neonatal samples.
Keratinocytes
Primary human keratinocyte cells from either adult or neonatal samples.
CD34+ cells
Pooled human hematopoietic progenitor cells derived from the umbilical cord blood
of mixed donors.
Neon Transfection System Cat. No. MPK5000
Shockingly simple transfection in stem cells
A next-generation electroporation technology for highly efficient delivery (~80%) of

nucleic acids such as episomal vectors into somatic cells.
Learn more at lifetechnologies.com/neon.
65
Detection
Validation is critical for your induced pluripotent stem cell (iPSC) research. Whether
you want to quickly confirm that cells are pluripotent or need flexibility in antibody
and dye choice, we have Molecular Probes labeling and detection assays and
imaging stations to help you get the confirmation you need. And, all of our labeling
and detection technologies are backed by our extensive internal and external support
resources, including the Molecular Probes online community and the trusted
Molecular Probes Handbook.
Learn more at lifetechnologies.com/detectpsc.
Alkaline Phosphatase Live Stain Cat. No. A14353
Alkaline Phosphatase (AP) Live Stain shows robust

staining in pluripotent stem cells. H9 human ESC
grown on MEF feeders show specific staining of the
pluripotent stem cell colonies.
Quick check for pluripotency
Rapidly detect pluripotency in expanding ESCs and iPSCs without compromising cell
integrity with Alkaline Phosphatase Live Stain.
Learn more at lifetechnologies.com/stemcellanalysis.
Antibodies
Flexibility and choice

Mix and match your choice of primary antibody and Molecular Probes fluorophore for
maximum flexibility in pluripotent stem cell imaging.
Learn more at lifetechnologies.com/antibodies.
H9 hESCs labeled with pluripotent surface antibodies:

SSEA4Alexa Fluor 488 (green), TRA-1-81Alexa
Fluor 594 (red), TRA-1-60Alexa Fluor 647 (purple).
FLoid Cell Imaging Station Cat. No. 4471136
Cell imaging right at your benchtop
Confidently detect and validate your ESC and iPSC colonies with
Live Alkaline Phosphatase and cell-surface antibodies on the
affordable, user-friendly FLoid Cell Imaging Station.
Learn more at lifetechnologies.com/floid.
66

Detection method selection guide.
Allows easy identification of

pluripotency
Allows easy identification of

pluripotency without compromising
cell integrity
Specific and flexible identification of

PSCs; antibodies can be combined
with different fluorophores for
co-staining
Product name
ELF 97 Endogenous Phosphatase

Detection Kit
Alkaline Phosphatase Live Stain
Primary antibodies
Cat. No.
E6601
A14353
Specificity
Medium (stains stem and progenitor

cells)
Medium (stains stem and progenitor cells)
High (stain human ESC and iPSC)
Terminal assay
Yes
No
No
Reaction time
Stains PSCs in 30 min
Stains PSCs typically in 20 min
Stain PSCs typically in 90120 min
Color/label
Yellow-green fluorescent precipitate

under Hoescht/DAPI longpass filter
Green/fluorescein (FITC)
Unconjugated primary antibodies,

may be detected with secondary
conjugated to fluorophore of choice
FLoid protocols
available
No
Yes
Yes
Unit size
500 L vial sufficient for staining four

24-well plates, twelve 6 cm dishes, or
four 10 cm dishes
50 L vial sufficient for staining four

24-well plates, twelve 6 cm dishes,
or four 10 cm dishes
100 g vial sufficient for staining four

24-well plates, twelve 6 cm dishes,
or four 10 cm dishes
Analyze
Assays
TaqMan Protein Assays

TaqMan Protein Expression Assays are designed for accurate quantitation of protein
expression using gold-standard TaqMan 5 nuclease chemistry with a workflow and
sample quantity similar to our all TaqMan Gene Expression and microRNA (miRNA)
Assays. As the protein expression results are obtained on the same analytical platform, these revolutionary assays enable direct correlation of mRNA and/or miRNA
expression to protein expression.
Learn more at lifetechnologies.com/taqmanprotein.
Selected instruments
Ion Proton Sequencer
Genome and exome sequencing on your benchtop.

Learn more at lifetechnologies.com/proton.
QuantStudio 12K Flex Real-Time PCR System
Delivering our highest throughput, flexibility, and scalability, the QuantStudio 12K
Flex Real-Time PCR System is an all-in-one qPCR instrument that features five interchangeable blocks to support 96-well, Fast 96-well, 384-well, TaqMan Array Card,
and OpenArray formats.
Learn more at lifetechnologies.com/quantstudio.
67
Mesenchymal stem cells (MSCs)

Culture
We provide the broadest selection of Gibco cGMP compliant complete culture systems for mesenchymal stem cells (MSCs) and
adipose-derived stem cells (ADSCs), some of which are free of animal-derived components. These media are designed to minimize
adaptation time, maximize cell performance, and help you meet regulatory requirements.
MSC culture system selection guide.

The standard serum-based
culture for MSCs and ADSCs
Reduced-serum medium
for lower variability and cost
savings with serum-culture
Fully documented serum-free MSC

culture designed to reduce the burden
in qualifying reagents and facilitate
transition from bench to clinic
Product
MSC-Qualified FBS
MesenPRO RS Medium
StemPro MSC SFM CTS
Cat. No.
12662002, 12662011, 12662029

(USDA), 12664025 (Australia
origin) 12763025 (US origin)
12746012
E071000 (New Zealand origin)
A1033201
Product use
IVD
IVD
Class II Medical Device
Recommended applications
Multiple species; culture of cells

sourced from bone marrow,
adipose tissue, cord blood,
umbilical cord, and placenta
Multiple species; culture

of cells sourced from bone
marrow, adipose tissue, and
umbilical cord
Human; culture of cells sourced from

bone marrow, adipose tissue, umbilical
cord, and hair follicle
High
Medium
Low
Components/unit
1 x 50 mL, 100 mL or 500 mL
1 x 500 mL MesenPRO RS
Basal Medium
1 x 10 mL MesenPRO RS
Growth Supplement
1 x 500 mL StemPro MSC SFM Basal

Medium
1 x 75 mL StemPro SFM Supplement
Additional components
required for complete
medium
DMEM, low glucose

(Cat. No. 11054020)
GlutaMAX-I
(Cat. No. 35050061)
GlutaMAX-I
(Cat. No. 35050061)
GlutaMAX-I CTS
(Cat. No. A1286001)
Substrate such as CELLstart CTS
(Cat. No. A1014201) or Attachment
Factor (Cat. No. S006100)
Supports growth at high

cell densityless medium,
surface area, and time
No
No
Yes
Stability of complete medium
Stable at 4C for 15 days
Specifically designed for cell

therapy research
No
No
Yes
MSC culture medium selection guide.

Serum-free, xeno-free MSC culture designed to reduce the risk and variability associated with animal-origin components
68
Product
StemPro MSC SFM XenoFree
Cat. No.
A1067501
Product use
RUO
Recommended applications
Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, and cord blood as well as
pericytes and fibroblasts
Low
Components/unit
1 x 500 mL StemPro MSC SFM Basal Medium

1 x 5 mL StemPro MSC SFM XenoFree supplement
Additional components
required for complete
medium
GlutaMAX-I CTS (Cat. No. A1286001)

Substrate such as CELLstart CTS (Cat. No. A1014201) or Attachment Factor (Cat. No. S006100)
Supports growth at high

cell densityless medium,
surface area, and time
Yes
Stability of complete medium

MesenPRO RS Medium Cat. No. 12746012
A reduced-serum medium for MSC culture containing 2% FBS, MesenPRO RS

Medium consistently improves expansion of MSCs compared with classical media
(DMEM + 10% FBS) and maintains trilineage mesoderm differentiation potential.
StemPro MSC Serum-Free Medium (SFM) Cat. No. A1033201
StemProMSC SFM provides superior efficiency of human MSC expansion at high

cell densities, requiring less medium, surface area, and time compared with classical
medium (DMEM (low glucose) + 10% FBS).
StemPro MSC SFM XenoFree Cat. No. A1067501
StemPro MSC serum-free, xeno-free medium offers a completely animal originfree

MSC culture system when used in conjunction with CELLstart CTS substrate.
MSCs and adipose-derived stem cells (ADSCs) are grown in a more physiologically
similar environment.
StemPro LipoMAX Supplement Cat. No. A1085001
StemPro LipoMAX supplement is a human-derived, lipoprotein-based cell culture

supplement for the improved expansion of MSCs and ADSCs.
CELLstart CTS Substrate Cat. No. A1014201
Xeno-free, fully defined cell culture substrate that contains components only of human
origin. CELLstart CTS substrate enables attachment of MSCs and ADSCs.
Cells
StemPro Human Adipose-Derived
Stem Cell Kit Cat. No. R7788110
Human ADSCs isolated from human lipoaspirate tissue and cryopreserved from
primary cultures. Kit contains human ADSCs and MesenPRO RS Medium.
Gibco Mouse (C57BL/6)

Mesenchymal Stem Cells Cat. No. S1502100
These MSCs are isolated from the bone marrow of a C57BL/6 mouse at 8 weeks
gestation.
StemPro Rat Alk Phos Expressing

Mesenchymal Stem Cells Cat. No. R7789120
These cells have a unique ability to track cells in transplantation and differentiation
studies.
Fluorescent visualization of StemPro Rat Alk Phos

Expressing Mesenchymal Stem Cells using the ELF
97 Endogenous Phosphatase Kit.
69
Differentiation
Gibco MSC differentiation kits
When used in conjunction with StemPro MSC SFM or MesenPRO RS Medium,

these kits provide a standardized culture workflow solution for MSC isolation, expansion, and differentiation into adipocytes, chondrocytes, and osteocytes.
Learn more at lifetechnologies.com/mscdiff.
StemPro Adipogenesis Differentiation Kit Cat. No. A1007001
Complete differentiation into fat cells
StemPro Chondrogenesis Differentiation Kit Cat. No. A1007101

Complete differentiation into cartilage cells
Mesenchymal stem cells differentiated into fat cells

and stained with Oil Red O, a fat-soluble dye.
StemPro Osteogenesis Differentiation Kit Cat. No. A1007201
Complete differentiation into bone cells
Analysis
The International Society for Cellular Therapy (ISCT) has proposed a set of standards to define hMSCs for laboratory investigations
and preclinical studies, including specific surface antigen expression in which 95% of the cells express the antigens recognized by
CD105, CD73, and CD90, with the same cells lacking (2% positive) the antigens recognized by CD45, CD34, CD14 or CD11b, CD79a
or CD19, and HLA-DR (Cytotherapy 8:315 (2006)).
Learn more about human MSC detection using the Attune Acoustic Focusing Cytometer at lifetechnologies.com/attune.
Human mesenchymal stem cells collected at passage 7 were identified as: (A) 99.7% CD73 +/0.29% CD19; (B) 99.4% CD73 +/0.22% CD45 ; (C) 99.7% CD90 +/0.09%
CD34; (D) 99.9% CD90 +/0.07% CD14; (E) 99.2% CD105 +/0.76% HLA-DR. The data demonstrate that, using Gibco mesenchymal stem cell media, cell expression
remains as expected after 7 passages.
70
Neural stem cells (NSCs)

Culture
Gibco media and supplements are the most widely cited reagents for use in neural stem cell culture. Choose between the flexibility of our B-27, N-2, StemPro Neural supplements and Neurobasal Medium or ease of use with the StemPro NSC SFM kit,
specifically formulated for serum-free growth and expansion of human neural stem cells.
StemPro NSC SFM Cat. No. A1050901
Designed for serum-free growth and expansion of human neural stem cells (hNSCs),
with versatility to support both adherent and suspension NSC cultures.
Neurobasal and Neurobasal-A Media
Serum-free media for the long-term viability of hippocampal and other neurons of the
central nervous system. Neurobasal Medium is optimized for prenatal and embryonic neurons, while Neurobasal-A medium is best for growing postnatal and adult
brain neurons.
Learn more at lifetechnologies.com/neurobasal.
Natural Mouse Laminin Cat. No. 23017015
Supports the growth of human or rat neural stem cells, as well as neural primary cells.
Human Neural Stem Cells (H9-Derived) Cat. No. N7800100

Consistent, high-purity cells with normal human karyotypes that can differentiate into
neurons, oligodendrocytes, and astrocytes. Consistent proliferation in adherent cell
culture when used with StemPro NSC SFM.
Rat Fetal Neural Stem Cells Cat. No. N7744100
These cells can be expanded as adherent or neurosphere suspension cell cultures

while more than 75% of NSCs retain undifferentiated phenotype. Also supports differentiation into neurons, astrocytes, and oligodendrocytes.
Phenotype marker expressions of Rat Fetal Neural
Stem Cells (proliferated up to passage 3) in StemPro
NSC SFM medium. Cells retain their undifferentiated
phenotype, and 90.9% cells express the NSC marker
nestin.
71
Differentiation
The following supplements are used in combination with a basal medium such as Neurobasal Medium.
B-27 Supplements
B-27 Supplements are 50X liquid supplements for growth and long-term viability
of hippocampal neurons. B-27 supplement is the most referenced supplement in
neurobiology research, and is offered in both serum-free and xeno-free versions.
Learn more at lifetechnologies.com/b27.
N-2 Supplement Cat. No. 17502048
N-2 Supplement is recommended for the growth and expression of neuroblastomas

as well as post-mitotic neurons in primary cultures from both the peripheral nervous
system (PNS) and the central nervous system (CNS).
G-5 Supplement Cat. No. 17503012

G-5 Supplement is a 100X supplement recommended for growth and expression of
glial cells (normal and tumor) of astrocytic phenotype (astrocytes).
Human embryonic stem cells differentiated into neurons.
72
Hematopoietic stem cells

(HSCs)
Hematopoietic stem cells expressing CD34 (CD34+ cells) are defined by their ability
to replenish all blood cell types and to self-renew. We offer CD34+ media, cells, and
selection tools specifically formulated to support the culture of these stem cells.
Culture
StemPro-34 SFM Cat. No. 10639011
Serum-free medium specifically formulated to support the development of human

hematopoietic stem cells (CD34+) from sources including bone marrow, peripheral
blood, and neonatal cord blood. It is manufactured with components of human,
recombinant, and synthetic origin.
StemPro CD34+ Cell Kit Cat. No. A14059
Pooled Human Hematopoietic Progenitor cells (HPCs) derived from the umbilical cord
blood pooled from multiple donors. Along with cord blood-derived CD34+ cells, this kit
includes StemPro-34 SFM basal liquid medium and frozen StemPro-34 Nutrient
Supplement to facilitate the immediate culturing of the StemPro CD34+ cells.
Dynal CD34 Progenitor Cell Selection System Cat. No. 11301D
Stem cells can be isolated directly from whole blood, cord blood, or bone using this
positive magnetic isolation of human CD34+ progenitor stem cells with bead release.
CD34+ cells, grown in StemPro-34 SFM, were reprogrammed into induced pluripotent stem cells using the
CytoTune-iPS Sendai Reprogramming Kit.
73
Cell analysis
Cell analysis
Life Technologies offers a diverse array of reagents and instruments for cell analysis researchDynabeads cell isolation technology, Molecular Probes fluorescent products for monitoring cell function and health, cell structure, and cell tracing, and a
broad portfolio of Molecular Probes and Novex primary and secondary antibodies for imaging and flow cytometry. We also offer
a selection of new instruments for flow cytometry (Attune Acoustic Focusing Cytometer and Tali Image-Based Cytometer) and
fluorescence imaging (FLoid Cell Imaging Station).
The chart below highlights the most popular products and new technologies in this chapter. Follow the Web links provided in this
If you have any questions about which of our solutions are right for you, please contact Life Technologies technical support by
phone, email, Facebook at facebook.com/cytometry, facebook.com/immunologynews, or facebook.com/LIFEMolecularProbes, or
Twitter at twitter.com/#!/MolProbes. Watch us on YouTube, too: youtube.com/lifetechnologies.
Section
Key technologies
Cell function and

health
Probes for visualizing

cell structure
Dynabeads cell isolation

technology, page 76
Click-iT EdU, page 82
CellLight fluorescent protein

constructs, page 87
Dynabeads CD3/CD28
technology, page 79
Premo FUCCI Cell Cycle

Sensor, page 83
CellROX Deep Red Reagent,
page 86
Cell isolation online selection

guide, page 76
LIVE/DEAD Viability/
Cytotoxicity Kits, page 81
Alexa Fluor phalloidins,

page 90
DynaMag magnets, page 80
CellEvent Caspase-3/7 Green

Detection Reagent, page 85
MitoTracker dyes, page 91
HulaMixer Sample Mixer,

page 80
pHrodo indicators, page 87
Cell analysis
Everyday essentials
Dynabeads cell
isolation technology
JC-1, page 91
LysoTracker dyes, page 92
BODIPY FL C5-ceramide,
page 93
TO-PRO-3 dye, page 94
Human cell isolation, page 77
Applications
Cell viability and proliferation,

page 81
Cytoskeleton, page 90
Mouse cell isolation, page 77

Positive, negative, and depletion
isolation strategies, page 78
Cell cycle, page 83

Apoptosis, page 84
Lysosomes and peroxisomes,

page 92
Mouse/human T cell activation

and expansion, page 79
Autophagy and reactive oxygen

species, page 86
ER and Golgi apparatus,

page 93
Endocytosis and phagocytosis,

page 87
Nucleus, page 94
Mitochondria, page 91
Intracellular ions, page 88
74
Cell analysis
HeLa cell labeled with CellLight Mitochondria-GFP (Cat. No. C10600) and
CellLight Talin-RFP (Cat. No. C10612) reagents and with Hoechst 33342
nucleic acid stain (Cat. No. H21492).
Probes for cell

tracing
Qtracker cell labeling,
page 95, 96
Instruments
Primary antibodies
for flow cytometry
Secondary antibodies
Attune Acoustic Focusing

Cytometer, page 100
Molecular Probes antibodies

for flow cytometry, page 102
Zenon antibody labeling,

page 108
Qdot nanocrystals for

multicolor flow cytometry,
page 105
APEX antibody labeling,

page 108
Attune Acoustic Focusing

Cytometer, page 100
Flow cytometer fluorophore

selection guide, page 104
FLoid Cell Imaging Station,

page 101
Primary antibody online

Alexa Fluor conjugated

secondary antibodies and
streptavidin, page 109
FluoroMyelin Green
Fluorescent Myelin Stain,
page 98
Calcein, AM, page 95, 96

CellTracker dyes, page 95
Fluorescent dextrans, page 95
Tali Image-Based Cytometer,

page 99
Neuronal tracing, page 97
Fluorescence SpectraViewer,
page 111
Fluorescence imaging,
page 101
Simplified multiplexing options,

page 103
Immunodetection strategies,
page 106
Flow cytometry, page 100, 102
Sample fixation and

permeabilization, page 105
Create your own labeled

antibodies, page 107
Image-based cytometry,
page 101
Cell analysis
Cell tracking, page 95
Secondary antibody online

Labeled secondary antibodies,

page 109
Choosing the right fluorophore,
page 110
Imaging tools, page 110111
75
Cell analysis
Dynabeads cell isolation technology

Gentle, reproducible separation of your targeted cells
Viable and functionalproducts for positive isolation, negative isolation, cell activation, and depletion
Gentlecolumn-free separation and inert bead surfaces

translate to gentler handling of your cells and reduce the
risk of contaminants in the preparations
High yieldstube-based separation allows you to achieve
excellent recovery of cells
Dynabeads are uniform spherical beads with highly defined and consistent product
characteristics, which helps ensure that you get truly reliable and reproducible
results.
Dynabeads FlowComp Mouse CD4
400
200
Viability:
86%
102
100
102
103
104
102
105
103
104
105
CD4+
CD4+
Column-based isolation
Purity:
78.5%
400
105
Propidium iodide
350
Count
300
250
200
150
100
104
103
102
Viability:
63%
50
0
Protocols for sample preparation and strategies for
102
103
104
102
105
103
104
105
CD4+
CD4+
1,000 1,250
Isolation of CD4+ T cells from mouse spleen cells. Cell isolation using Dynabeads
FlowComp Mouse CD4 results in substantially higher purity (97%) and viability
(86%) than column-based positive cell isolation (yielding purity and viability of 78%
and 63%, respectively).
250
500
500
Count
1,000
Count
750
cell isolation
Data showing the performance of our cell isolation products
versus other commercially available products
Help in choosing the correct magnets for your
tubes or plates
Links to videos, brochures, and application notes and to
references that cite the use of Dynabeads
103
2,000
isolation product
300
104
1,500
Cell isolation and activation
Selection guides for choosing the correct cell
105
Propidium iodide
500
The table below organizes our cell isolation products according

to cell origin and isolation method. If you go to lifetechnologies
.com/cellisolation you can find information about our entire line
of cell isolation products, and youll also have access to:
Purity:
97.2%
600
Count
When cells are removed from their natural environment, there

is a risk that experimental procedures will negatively impact
cell phenotype and function. Choosing the right cell separation
method is therefore critical to your downstream experiments.
Dynabeads are superparamagnetic, monosized polymer
beads coated with a thin, inert polymer shell to encase the
magnetic material. This design helps to eliminate the risk that
any unwanted material such as iron is left in the sample after
separation. In addition, since the cells are never touched by the
beads (negative-isolation and depletion beads) or are released
from the beads after the gentle magnetic separation (positive-isolation beads), you get high purity and viability with no
process-derived remnants that could affect your downstream
results.
102
103
104
CD4 FITC
105
102
103
104
105
CD4 FITC
Purity of human CD4+ T cells. Purity before (left) and after (right) negative isolation
from PBMC using Dynabeads Untouched Human CD4 T Cells.
76
Cell analysis
Human cell isolation
Gentle tube-based isolation of human cells directly from whole blood, MNC, buffy coat,
bone marrow, or tissue samples for any downstream assay, including flow cytometry.
Dynabeads are available for isolating human T cells, B cells, stem cells, NK cells,
monocytes, dendritic cells, endothelial cells, tumor cells, leukocytes, granulocytes.
Incubate your
starting sample
with Dynabeads
Learn more at lifetechnologies.com/humancellisolation.

Dynabeads are available for:
Positive isolation and cell release

Negative isolation resulting in untouched cells
Depletion of unwanted cell types or positive cell isolation for
Separate
bead-captured
cells with a
Dynal MPC
magnet
molecular applications
Isolate cells using your own antibody

If you cant find a ready-to-use product for human cell isolation, we have a range of
Dynabeads products that can be combined with an antibody of your choice to create
a tailored cell isolation tool: Streptavidin Dynabeads, Secondary-coated Dynabeads,
and Surface-activated Dynabeads.
Learn more at lifetechnologies.com/humancellisolation.
Transfer
supernatant with
untouched cells
to a new tube
Mouse cell isolation
Gentle tube-based isolation of mouse cells from directly from whole blood, spleen, lymph
node, or thymus for any downstream assay, including flow cytometry. Dynabeads are
available for isolating mouse T cells, B cells, NK cells, and dendritic cells.
Learn more at lifetechnologies.com/mousecellisolation.
Dynabeads are available for:
Positive isolation and cell release

Negative isolation of untouched cells
Depletion of unwanted cell types or positive cell isolation for
molecular applications
If you cant find a ready-to-use product for human cell isolation, we have a range of
Dynabeads products that can be combined with an antibody of your choice to create
a tailored cell isolation tool: Streptavidin Dynabeads, Secondary coated Dynabeads,
and Surface activated Dynabeads.
Positively
isolated cells
(bead-captured)
Cell-based assays, flow

cytometry, cell expansion
eads
ove b
Rem
Molecular applications
e.g., mRNA and DNA analysis
Isolate cells using your own antibody
Negatively
isolated cells
(untouched)
1
Workflow diagram for using Dynabeads for positive

or negative tube-based cell isolation.
Learn more at lifetechnologies.com/mousecellisolation.
77
Cell analysis
Positive isolation
Positive-isolation kits use Dynabeads to isolate cells of interest directly from all types
of sample material (e.g., human or mouse whole blood, bone marrow or peripheral
blood mononuclear cells, spleen, or lymph nodes) by binding to antibody-coated Dynabeads. Positive isolation is ideal when cells of the highest purity and viability are
required. Cells may be used for downstream analysis or as source material for the
purification of RNA or proteins from the isolated fraction.
FlowComp kits give you positive isolation of pure and viable cells directly from whole
blood/buffy coat or PBMC. And, after a simple step in which the beads are removed,
the cells can be used directly in all downstream experiments, including flow cytometry.
Positive isolation:
Ideal for flow analysis and cell-based assays

When maximum purity and yield is critical
From whole blood, bone marrow, or buffy coat
Learn more at lifetechnologies.com/cellisolation.
Negative isolation
Negative isolation (Untouched) kits use Dynabeads in combination with an antibody

cocktail to remove all unwanted cells from your sample, leaving only the target cells
of interest behind. These kits are optimal for the isolation of fragile cells or when
there are concerns that activation via antibody binding can interfere with downstream
assays. Our antibody cocktails have been optimized to give you high cell purity and
yield, and your resulting cells are both antibody- and bead-free.
Negative isolation:
Truly untouched cells, bead- and antibody-free

Cells in perfect shape for any application
High yield, purity, and viability
Depletion
Dynabeads depletion products are ideal for removal of one unwanted cell type at a
time from your sample. Dynabeads are precoated with an antibody toward one target
cell type that can be depleted from all types of starting materialeven viscous materials such as whole blood.
Depletion:
Easy removal of unwanted cell types

Ideal for positive cell isolation for molecular applications
High yield and purity
78
Cell analysis
Dynabeads for mouse and human T cell

activation and expansion
Mimic in vivo activation of T cells
No need for antigen-presenting cells (APC) or antigen

Gentle and easy, yet efficient, robust, and highly reproducible
Activated cells retain their functional properties
Use the same technology for mouse and human research-grade studies and
Nature
Dendritic
cell
Resting T cell
human clinical research

The activation of naive and memory T cells is initiated through contact dependent
antigen recognition involving antigen-presenting cells (APCs) such as dendritic cells.
Activation occurs through the engagement of the T cell receptor complex (TCR/CD3,
signal 1) and the CD28 costimulatory molecule (signal 2) on the surface of the T cell.
Both signals are required to trigger cell-mediated immunity. Triggering of signal
1 alone can lead to T cell inactivation by anergy or apoptosis. Dynabeads T cell
activation and expansion products offer a simple solution for mimicking the in vivo
interaction of T cells with APCs by utilizing the two activation signals, and a threedimensional bead similar in size to APCs. Dynabeads CD3/CD28 can be used for both
short-term activation and long-term expansion and are easily removed from culture.
Activated cells are bead- and antibody-free, and their functions, cytokine profiles, and
T cell repertoires reflect those of in vivo activation.
Activated T cell
CD28
CD3/TCR
Expansion platform
Resting T cell
Anti-CD28
Bead
Activated T cell
CD28
CD3/TCR
Anti-CD3
The Dynabeads T cell activation/expansion platform

mimics in vivo activation/expansion.
Dynabeads CD3/CD28 technology is the only activation solution that allows for a
controllable setting. Dynabeads CD3/CD28 technology is designed with covalently
linked anti-CD3 and anti-CD28 antibodies for simple use and easy removal. Several
CD3/CD28 antibody clones and ratios are available for more flexibility and optimization
for your particular application. Custom conjugations are also available.
Learn more about Dynabeads for mouse and human T cell activation
at lifetechnologies.com/cellactivation.
2970
3,000
Fold expansion
Dynabeads CD3/CD28/CD137 is specifically optimized for activation and expansion of

mouse and human antigen-specific T cells. In addition to the anti-CD3 and anti-CD28
antibodies, the beads are also coated with anti-CD137 that recognizes CD137 (4-1BB),
a costimulatory molecule expressed primarily but not exclusively on activated CD8+
and CD4+ T cells. Thus, Dynabeads precoated with monoclonal antibodies towards
CD3, CD28, and CD137 specifically enhance proliferation of antigen-specific T cells.
3,500
2,500
2,000
Dynabeads CD3/CD28/CD137 for antigen-specific T cell

activation and expansion
1615
1,500
865
1,000
446
500
0
2.4
4.7
10.9
20.5
50
190
10
11
12
Days
Expansion of naive T cells from human peripheral

blood using Dynabeads Human T-Activator CD3/
CD28 (Cat. No. 11131D) for 12 days.
79
Cell analysis
Accessories for Dynabeads products
Get the highest utility and efficiency from Dynabeads products

DynaMag magnets
DynaMag magnets are optimized for efficient cell isolation using Dynabeads. The
DynaMag magnets combine a strong magnetic attraction for efficient separations
with flexible and smart ergonomic designs that ensure comfortable working positions. The magnets can be used for any starting sample in combination with all types
of Dynabeads.
We offer DynaMag magnets for these tube and plate configurations:
1.52 mL microcentrifuge tubes (straight tube racks and circular racks)

(Cat. Nos. 12321D, 12320D)
0.55 mL, 15 mL, and 50 mL tubes (Cat. Nos. 12303D, 12301D, 12302D)
6-well plates
24-well plates
96-well plates (side magnet and bottom magnet versions, skirted and half-skirted
DynaMag-5 Magnet (Cat. No. 12303D)
plate versions, round- and flat-bottomed)
PCR strips (8 and 12 wells)

48- and 96-microtubes for the GeneAmp PCR System
Closed, sterile blood bags (50330 mL static separations and >10 L in continuous
flow separations)
Find specifications for all of these magnets online at lifetechnologies.com/magnets.
HulaMixer Sample Mixer
The HulaMixer Sample Mixer is perfect for sample preparation with Dynabeads
products and for any other application that requires thorough mixing.
The HulaMixer offers the following features:
Tilt, rotate, and/or vibrate your samples

Continuous or timed operation
All settings easily adjustable
For use at 4C to 40C
Two different platforms supplied to suit all kinds of tubes (0.550 mL)
See the HulaMixer Sample Mixer in action at lifetechnologies.com/hulamixer.
HulaMixer Sample Mixer (Cat. No. 15920D).
80
Cell analysis
Cell viability, proliferation,

and health
Lymphocyte gate
Life Technologies offers a diverse selection of reagents for assessing cellular function and health. Many of the assays are fluorescence-based or colorimetric, offering
sensitivity and convenience as well as safety. These products have been validated
on multiple instrument platforms, including microscopy, flow cytometry, microplate
readers, and high-content screening, enabling the analysis of a broad spectrum of
cell functions:
Viability and cytotoxicity

Proliferation
Apoptosis
Autophagy
Oxidative stress
Endocytosis and phagocytosis
Intracellular ions
SSC
Viability gate
B
Live cells
Dead cells
0.55%
Learn more about all of our cell function assays at lifetechnologies.com/cellfunction.
Viability and cytotoxicity
Fixable Violet
CD4 Cy55PE
Fluorometric assays of cell viability and cytotoxicity are easy to perform with the use
of a fluorescence microscope, fluorometer, fluorescence microplate reader, or flow
cytometer, and they offer many advantages over traditional colorimetric and radioactivity-based assays. Our viability and cytotoxicity assay reagents and kits are principally used to enumerate the proportion of live and dead cells in a population.
LIVE/DEAD Viability/Cytotoxicity Kits
Simple to performadd, incubate, and analyze; no wash steps required

Versatilecompatible with adherent cells, nonadherent cells, and certain tissues
Simple to analyzegreen-fluorescent cells are live, red-fluorescent cells are dead
CD4 Cy55PE
Learn more at lifetechnologies.com/livedeadkits.
CD8 Q705
Cell viability and health
The LIVE/DEAD Viability/Cytotoxicity Kits provide an exceptionally easy fluorescencebased method for determining viability of adherent or nonadherent cells and for
assaying cytotoxicity. LIVE/DEAD assays typically employ two separate fluorescence
probes for clear live/dead distinction, and we have developed kits in several colors for
flow cytometry (see right), microscopy (see below), or microplate formats.
CD8 Q705
Fluorescence staining of live and dead cells with the LIVE/DEAD Cell Imaging Kit. HepG2 cells grown in 96-well
microplate wells were stained with the LIVE/DEAD Cell Imaging Kit (Cat. No. R37601) and imaged on a Nikon
Eclipse T200 microscope. Live cells are stained green; dead cells are stained red.
Live-cell gating with the LIVE/DEAD Fixable Violet

Dead Cell Stain Kit (Cat. No. L34955) minimizes
staining artifacts from analysis. In a comparison
between live-cell gating using scatter (A), and livecell gating using LIVE/DEAD Fixable Violet dye (B),
staining artifacts with scatter gating are illustrated.
The significant number of dead cells in subsequent
analysis using scatter (C) are noted, as compared to
the use of the LIVE/DEAD Fixable Violet dye (D) to
eliminate dead cells. Reproduced with permission
from Elsevier (J Immunol Methods 313:199(2006)).
81
Cell analysis
SYTOX Dead Cell Stains
capabilities
Ease of use: no wash steps; simply add, incubate, and analyze
Fixable: Staining retained after fixation for simple live/dead analysis with intracellular phenotyping
The cell-impermeant nucleic acid SYTOX Dead Cell Stains easily penetrate eukaryotic cells and both gram-positive and gram-negative bacteria with compromised
plasma membranes, yet are completely excluded from live cells. Because they do
not cross intact cell membranes and increase fluorescence upon double-stranded
DNA binding, they are among our most brilliant dead cell stains. Easy to use, SYTOX
dead cell stains can be applied to cells and visualized without an additional wash step
because they are nonfluorescent in aqueous media. These stains can be used with
multiple platforms including fluorescence microscopy, flow cytometry (see right), and
microplates.
See all colors of SYTOX Dead Cell Stains at lifetechnologies.com/sytox.
1,000
800
Number of cells
Accuracy: high-affinity nucleic acid stains for easy dead-cell discrimination

Flexibility: multiple colors with minimal spectral overlap for expanded multicolor
600
400
200
0
101
102
103
104
105
106
SYTOX Green fluorescence
Dead-cell discrimination using SYTOX Green Dead

Cell Stain. A mixture of heat-killed and live Jurkat cells
was stained with 30 nM SYTOX Green Dead Cell Stain
(Cat. No. S34860) at room temperature for 20 min.
Cells were analyzed on the Attune Acoustic Focusing
Cytometer equipped with a 488 nm laser. Fluorescence
emission was collected using a 530/30 nm bandpass
filter. Live cells are easily distinguished from the
brighter dead-cell population.
Cell proliferation and cell division
Cell proliferation and the characterization of agents that either promote or retard proliferation are extremely important areas of cell biology and drug-discovery research. We
offer both traditional reagents for assessing cell proliferation and the cell cyclein
particular the Hoechst nucleic acid stains and probes for 5-bromo-2-deoxyuridine
(BrdU) incorporation during cell divisionas well as more advanced tools developed
in our laboratories, including the Click-iT EdU cell proliferation assay.
Click-iT EdU Cell Proliferation Assays
Accurate: superior to BrdU assays with minimal variation (low CVs) and much
simpler protocol
Speed: results in as little as 90 min

Simplicity: streamlined protocol with fewer components than the original kits
Versatility: assays for flow cytometry, imaging, HTS, HCS, and whole-animal
interrogations
Click-iT EdU cell proliferation assay utilizes the power of click chemistry to provide
a superior alternative to traditional methods for detecting and quantitating newly
synthesized DNA. Click-iT assays use a modified nucleoside, EdU (5-ethynyl-2deoxyuridine), that is incorporated during DNA synthesis in a quick click chemistry
reaction. The Click-iT EdU assay protocol is compatible with both adherent cells and
cell suspensions. From start to finish, the EdU detection assay is complete in as little
as 90 minutes, as compared with the antibody-based BrdU method, which takes 624
hours to complete. In addition, the Click-iT EdU cell proliferation assay can be multiplexed with surface and intracellular marker detection using Alexa Fluor dyelabeled
secondary antibodies. Although the majority of applications are in cultured mammalian cells, Click-iT EdU reagents and methods have also been successfully applied to
a wide range of model organisms.
See all of the Click-iT assays at lifetechnologies.com/clickit.
82
Comparison of EdU and BrdU signal brightness.

Sectioned formaldehyde-fixed, paraffin-embedded
tissue was prepared from rats injected intraperitoneally with BrdU (upper panel) or EdU (lower panel).
Proliferating cells within the intestinal villi that
incorporated the nucleoside are pink. Nuclei in both
samples were counterstained with Hoechst 33342
(gray). Images were acquired with a 20x objective
on a Nikon Eclipse 800 epifluorescence microscope.
Cell analysis
Superior performancebright single-peak staining enables visualization of
multiple generations
Long-term signal stabilitywell retained in cells for several days post-staining

Noncytotoxicno known effect on proliferative ability or biology of cells
Versatilemultiple colors available to easily combine with antibodies or markers
of cell function, such as GFP
CellTrace reagents allow you to permanently label cells to trace generations or divisions in vivo or in vitro without affecting morphology or physiology. We offer CellTrace
reagents that are compatible with violet laser, 488 nm laser, and 633/635 nm laser
excitation on your flow cytometer.
See all of our CellTrace reagents at lifetechnologies.com/celltraceproliferation.
Premo FUCCI Cell Cycle Sensor
Live-cell indicator of cell cycle status

Flexible: works with live cells, but compatible with fixation
Convenient: add to cells, incubate overnight, and visualize
The fluorescence ubiquitination cell cycle indicator (FUCCI) is a genetically encoded,
two-color (red and green) indicator that allows you to follow cell division within a cell
population (see below). We have incorporated the FUCCI genetic constructs into the
powerful BacMam gene delivery systemthe Premo FUCCI Cell Cycle Sensor
allowing a simple and efficient method for labeling cells and following their division.
Number of cells counted
CellTrace reagents for flow cytometry cell proliferation studies
100
101
102
103
104
CellTrace CFSE fluorescence

Following cell proliferation in human peripheral
blood lymphocytes using the CellTrace CFSE Cell
Proliferation Kit (Cat. No. C34554). Human peripheral blood lymphocytes were harvested and stained
with CellTrace CFSE (carboxyfluorescein diacetate,
succinimidyl ester; 5(6)-CFDA, SE) on day 0. A portion
of the population was arrested at the parent generation
using mitomycin C (red peak). The remainder of the
sample was stimulated with phytohemagglutinin and
allowed to proliferate for 5 days. Solid green peaks
represent successive generations.
The Premo FUCCI Cell Cycle Sensor can be used to assess the effect of drugs,
siRNA, or other factors on the transition of cells through the cell cycle. The fluorescence signals from geminin-GFP and Cdt1-RFP have been demonstrated to be resistant to fixation with 4% formaldehyde and permeabilization with 0.1% Triton X-100,
thereby enabling processing of labeled cells with antibodies to other cellular targets.
Learn more about the Premo FUCCI Cell Cycle Sensor at lifetechnologies.com/
fucci.
M
Co
lor
le
s
P)
ed
(C
d t1
-RF
P)
Green
(ge
mi
nin
-G
F
S
Yellow
Imaging cell cycle progression in live cells with the Premo FUCCI Cell Cycle Sensor.(A) Schematic of cell cycle
progression with nuclear fluorescence changes. (B) U2OS cells were transduced with the Premo FUCCI Cell
Cycle Sensor (Cat. No. P36237), then stained with Alexa Fluor 647 wheat germ agglutinin (Cat. No. W32466).
83
Cell analysis
Apoptosis
Apoptosis (programmed cell death) is the genetically controlled ablation of cells during normal development.
Inappropriately regulated apoptosis is implicated in disease states such as Alzheimers disease, stroke, and
cancer. In contrast to necrotic cells, apoptotic cells are characterized morphologically by compaction of
the nuclear chromatin, shrinkage of the cytoplasm and production of membrane-bound apoptotic bodies.
Biochemically, apoptosis is distinguished by fragmentation of the genome and cleavage or degradation of
several cellular proteins.
As with cell viability, no single parameter fully defines cell death in all systems; therefore, it is often advantageous to use several different approaches when studying apoptosis. Anticancer drug candidates failing to
induce apoptosis are likely to have decreased clinical efficacy, making apoptosis assays important tools for
high-throughput drug screening.
Membrane asymmetry probe
Accurate apoptotic analysis on trypsinized cells

Simple 5-minute staining protocol
Compatible with other blue-excited apoptotic stains
The Violet Ratiometric Membrane Asymmetry Probe/Dead Cell Apoptosis Kit (Cat. No. A35137) provides an
easy, efficient method for the detection of apoptosis with dead-cell discrimination using a violet laser flow
cytometer (see figure, below). The Violet Ratiometric Membrane Asymmetry Probe detects the membrane
asymmetry changes during apoptosis. It works well on adherent and suspension cells and correlates with
other indicators of apoptosis, such as caspase detection and changes in mitochondrial membrane potential.
The dye exhibits an excited-state intramolecular proton transfer (ESIPT) reaction resulting in dual fluorescence with two emission bands corresponding to 530 nm and 585 nm, producing a two-color ratiometric
response to variations in surface charge. The F2N12S probe is combined with SYTOX AADvanced dead cell
stain, which is capable of passing through the cell membrane only in late apoptotic or necrotic cells, allowing
discrimination from early apoptotic cells.
262,144
196,608
A+D
131,072
65,536
L
65,536
131,072
196,608
262,144
F2N12S green fluorescence
F2N12S green fluorescence
Unlike annexin-based assays, this assay does not require special buffers or wash steps, and it is less susceptible to the cell membrane damage commonly found during the physical or chemical removal steps when
assaying adherent cells, therefore providing better data quality.
196,608
A+D
131,072
65,536
L
262,144
65,536
105
D
104
103
102
101
1,250
2,500
3,750
F2N12S ratio 585 nm/530 nm

fluorescence x 1,000
84
131,072
196,608
262,144
F2N12S orange fluorescence
5,000
SYTOX AADvanced fluorescence
SYTOX AADvanced fluorescence
F2N12S orange fluorescence
105
D
Violet ratiometric membrane asymmetry probe for

apoptosis detection. Jurkat cells (T-cell leukemia,
human) were treated with 10 M camptothecin for
four hours (panels B and D) or left untreated as a
control (panels A and C). Samples were analyzed
on a flow cytometer with 405 nm excitation using
585 nm and 530 nm bandpass filters for F2N12S,
and 488 nm excitation for SYTOX AADvanced dead
cell stain using a 695 nm bandpass filter. Live cells
can be discriminated from apoptotic and dead cells
by the relative intensities of the two emission bands
from F2N12S (A and B). In panels C and D, SYTOX
AADvanced dead cell stain fluorescence is plotted
against a derived ratio parameter from the two emission bands (585/530 nm) of F2N12S. A = apoptotic cells,
L = live cells, D = dead cells.
104
103
L
A
102
101
1,250
2,500
3,750
5,000
F2N12S ratio 585 nm/530 nm

fluorescence x 1,000
Cell analysis
CellEvent Caspase-3/7 Green Detection Reagent
Optimized caspase-3/7 substrate for apoptosis analysis

Simple, no-wash protocol helps preserve delicate apoptotic cells
Compatible with both live-cell fluorescence imaging and formaldehyde-based fixation methods
The CellEvent Caspase-3/7 Green Detection Reagent (Cat. No. C10423) is a novel
fluorogenic substrate for activated caspase-3/7 that is compatible with both live-cell
and fixed-cell imaging, with absorption/emission maxima of ~502/530 nm. Activation
of caspase-3 is considered an essential event during apoptosis, making this an optimized reagent for analysis of apoptotic cells.
The CellEvent Caspase-3/7 Green Detection Reagent is a fouramino acid peptide
(DEVD) conjugated to a nucleic acid binding dye with an absorption/emission maxima
of ~502/530 nm. The DEVD peptide sequence is a cleavage site for caspase-3/7, and
the conjugated dye is nonfluorescent until cleaved from the peptide and bound to DNA.
The CellEvent reagent is intrinsically nonfluorescent as the DEVD peptide inhibits the
ability of the dye to bind to DNA. However, after activation of caspase-3/7 in apoptotic
cells, the DEVD peptide is cleaved, enabling the dye to bind to DNA and produce a
bright fluorogenic response. The fluorescence emission of the dye when bound to
DNA is ~530 nm and can be observed using a standard FITC filter set. CellEvent
reagent is ideal for imaging (see right), high-content screening, and flow cytometry.
Multiplex imaging of apoptosis. U2OS cells were

treated with 30 M etoposide for 18 hr to induce apoptosis. The treated cells were stained first with 7.5 M
CellEvent Caspase-3/7 Green detection reagent (Cat.
No. C10423, green fluorescence) to detect apoptosis
and Hoechst 33342 nucleic acid stain (Cat. No. H3570,
blue fluorescence) to label nuclei, and then with
150 nM MitoTracker Deep Red FM (Cat. No. M22426,
pink fluorescence) to label mitochondria. Following
fixation and permeabilization, actin was labeled with
Alexa Fluor 546 phalloidin (Cat. No. A22283, orange
fluorescence).
Click-iT TUNEL Alexa Fluor Imaging Assay
Robust workflow can be completed typically in 3 hours

Available with Alexa Fluor 488, 594, and 647 dyes
Much more sensitive than traditional antibody-based TUNEL methods
Multiplexable with most other fluorescent reagents
Ideal for imaging in vivo and in vitro, compatible with HCS and microplate assays
See all of the Click-iT assays at lifetechnologies.com/clickit.
The Click-iT TUNEL assay (see right) is a breakthrough technology based on click
chemistry for detection of DNA fragmentation. For TUNEL assays to yield meaningful
results, it is necessary not only that the modified nucleotide is an acceptable substrate
for TdT, but also that the detection method is sensitive and avoids detrimental loss of
cells from the sample.
Late-stage apoptosis visualized using the Click-iT

TUNEL Imaging Assay. HeLa cells were treated with
staurosporine, then fixed and permeabilized. The
Click-iT TUNEL Alexa Fluor 647 Imaging Assay (Cat.
No. C10247) was performed. Activated caspase-3 was
detected with a rabbit polyclonal primary antibody
for cleaved caspase-3 and labeled with Alexa Fluor
488 goat antirabbit IgG antibody (green). Tubulin
was detected with a mouse monoclonal anti-tubulin
antibody and labeled with Alexa Fluor 555 goat
antimouse IgG (orange). Nuclei were stained with
Hoechst 33342 (blue). The light blue color represents
an overlay of caspase (green), Hoechst (blue), and
TUNEL (magenta) signals.
85
Cell analysis
Autophagy
Autophagy describes the segregation and delivery of cytoplasmic cargo, including

proteins and organelles, for degradation by hydrolytic enzymes. The LC3B protein can
be used as a general marker for autophagic membranes, and we developed the products below for imaging the LC3B protein and autophagic organelles in live cells.
Premo Autophagy Sensors

The Premo Autophagy Sensors (Cat. Nos. P36235 and P36236, see right) combine
the selectivity of an LC3B-fluorescent protein chimera (GFP or RFP) with the transduction efficiency of BacMam technology, enabling unambiguous visualization of this
protein in live cells. The reagent also tolerates fixation with formaldehyde and thus is
compatible with fixed-cell analysis, which is required for multiplex experiments using
antibodies, and is preferred for HCS analysis workflows. To image autophagy, simply
add BacMam LC3-FP to cells and incubate overnight for protein expression.
LC3B Antibody Kit

The LC3 Antibody Kit for Autophagy (Cat. No. L10382) includes a highly specific
rabbit polyclonal antibody against LC3B that has been validated for use in fluorescence microscopy and high-content imaging. Also included is a control compound for
inducing autophagosomesfollowing treatment with this compound, lysosomal pH
increases and the normal autophagic flux is disrupted, resulting in autophagosome
accumulation.
Learn more about LC3B protein detection at lifetechnologies.com/autophagy.
Expression of the Premo Autophagy Sensor LC3BGFP. (A) Imaging of Premo Autophagy Sensor in
rat hippocampal neurons. (B) Imaging of Premo
Autophagy Sensor (green) and CellLight LysosomesRFP (red) in HeLa cells.
Oxidative stress
CellROX Deep Red Reagent
Compatible with fluorescent imaging, florescent plate readers, flow cytometry, and
high-content screening
Highly resistant to photobleaching

Multiplexible: compatible with GFP and Alexa Fluor 488 dye
Versatile: suitable for live-cell imaging or fixed samples
CellROX oxidative stress reagentsincluding CellROX Green reagent (Cat. No.

C10444), CellROX Orange reagent (Cat. No. C10443), CellROX Deep Red reagent
(Cat. No. C10422), and a variety pack (Cat. No. C10448)are fluorogenic probes
designed to reliably measure reactive oxygen species (ROS) in live cells. The cellpermeable CellROX reagents become brightly fluorescent upon oxidation, with
excitation/emission maxima at 485/520 nm, 545/565 nm and 640/665 nm, respectively.
The CellROX Green reagent binds to DNA (fluorescent signal is localized primarily in
the nucleus and mitochondria). In contrast, CellROX Orange and Deep Red reagents
localize to the cytoplasm.
The resulting fluorescence can be measured using traditional fluorescence
microscopy, high-content imaging and analysis, microplate fluorometry, or flow
cytometry. The staining workflow is simple, and the reagent can be applied to
cells in complete growth medium or buffer. All of the CellROX reagents are very
photostable when compared with traditional ROS detection dyes. In addition,
both CellROX Green and CellROX Deep Red reagents are retained in cells after
formaldehyde fixation, providing assay flexibility and improved workflows when
compared with classic dyes used for ROS detection. CellROX Green staining is also
stable to detergent permeabilization.
86
Imaging oxidative stress with CellROX Deep Red

reagent. (A) Human osteosarcoma (U2OS) cells were
treated with 100 M menadione for 1 hr to induce
oxidative stress, and then stained with 5 M CellROX
Deep Red reagent, 5 g/mL of CellMask Orange
plasma membrane stain, and 1 M SYTO Green
fluorescent nuclear stain for 30 min at 37C. The
cells were washed 3 times with HBSS before imaging.
Cell analysis
Endocytosis and phagocytosis
The pHrodo dye and CellLight reagents are fluorescence-based tools that can be
used to interrogate routes of molecules and microbeswhether critical to cell health
or pathogenicacross the plasma membrane, as well as their final destination after
entering the cell.
pHrodo indicators
The proprietary pH-sensitive pHrodo dye is a specific sensor of endocytosis. The dye
is nonfluorescent at neutral pH and fluoresces bright red in acidic environments (see
right). This increase in fluorescence signal at low pH makes it ideal for studying endocytosis and its regulation by drugs and/or environmental factors. For multiplexing,
pHrodo dye is available in red and green colors, and the signal can be preserved by
formaldehyde fixative. pHrodo dye is available as convenient conjugates of E. coli,
S. aureus, and Zymosan A BioParticles for phagocytosis and dextran BioParticles
for endocytosis. If you want to make your own conjugations, both pHrodo green and
red are available in various reactive chemistries and in antibody labeling kits. The
pHrodo dye allows you to:
Discriminate endocytosis from adherent and extracellular particles

Specifically detect phagocytosis and endocytosis with a fluorogenic dye
Reduce signal variability and improve timing in sensitive experiments
Can be multiplexed with other fluorescent dyes
Learn more about all of our pHrodo indicators at lifetechnologies.com/phrodo.
CellLight reagents
CellLight reagents label a number of intracellular organelles (see below), including
endosomes, with either GFP or RFP. CellLight reagents use fluorescent proteins that
survive fixation and permeabilization, and therefore they can be used in combination
with any antibody. Labeling of transferrin, epidermal growth factor, or low-density
lipoprotein with Alexa Fluor dyes provides a useful means of visualizing ligand internalization. Some of our more popular fluorescent probes are also available in 0.1 mL
format, providing a cost-effective way for researchers to try these innovative products.
Human osteosarcoma (U2OS) cells expressing CellLight Tubulin-GFP (Cat. No. C10509) were treated
with 200 M tert-Butyl hydroperoxide for 2 hr. A stain
solution containing 5 M CellROX Orange (Cat. No.
C10443), and 2 drops/ml of NucBlue Live Cell Stain
(Cat. No. R37605) was applied for 30 min at 37C.
Cells were washed and imaged with Live Cell Imaging
Solution (Cat. No. A14291DJ).
Time-lapse images showing internalization and acidification of the pHrodo E.coli BioParticles conjugate
during phagocytosis by Murine J774A.1 cells. Using
excitation with a 561 nm laser, fluorescence images
were recorded in the 570699 nm emission range and
white-light DIC images were recorded on a separate
PMT. Images were collected every 30 sec for 83.8 min
(only the first and last images in the time course are
shown here) using a Leica TCS SP5 laser confocal
microscope employing a 63 (NA 1.4) oil objective and
the resonant galvanometer scanner mode (8000 Hz).
Image contributed by Lucy Deriy and Deborah Nelson,
University of Chicago. See Nelson Lab time-lapse
video of this process.
Human osteosarcoma (U2OS) cells expressing

CellLight Actin-RFP were treated with 100 M
menadione for 1 hr. A stain solution containing 5 M
CellROX Green (Cat. No. C10444) and 2 drops/mL
of NucBlue Live Cell Stain (Cat. No. R37605) was
applied for 30 min at 37C. Cells were washed and
imaged with Live Cell Imaging Solution (Cat. No.
A14291DJ).
87
Cell analysis
Intracellular ions
Ion channels provide exciting opportunities for drug development, targeting a diverse
array of therapeutic areas including neurological, cardiovascular, and pulmonary
diseases, pain, and inflammation. To address the most challenging ion channel targets
in any desired cell background, researchers need an assortment of optimized assays
for their discovery efforts. Our product offering for cell-based ion channel assays is
designed for sensitivity, ease-of-use, and assay flexibility.
Learn more at lifetechnologies.com/ionchannelassays.
Cell-based ion channel assays.
Ion
Assay technology
Description
Potassium
FluxOR Potassium
Ion Channel Assay
Functional, HTS-validated universal potassium

assay (Cat. Nos. F10016, F10017)
lifetechnologies.com/fluxor
BacMam Ion Channel
Targets
BacMam-mediated delivery of potassium ion

channels including hERG
lifetechnologies.com/bacmamionchannels
Calcium
Fluo-4 Direct
Calcium Assay Kit
Add-and-read, homogenous fluorescent

calcium readout validated for HTS
(Cat. Nos. F10471, F10472, F01473)
lifetechnologies.com/fluo4direct
Premo Cameleon
Calcium Sensor
Fluorescent proteinbased calcium biosensor

protein
lifetechnologies.com/premocameleon
Chloride
Premo Halide
Sensor
Membrane
potential
Voltage sensor probes
Fluorescent proteinbased chloride biosensor

protein
lifetechnologies.com/premohalide
FRET-based, ratiometric assessment of
membrane potential
lifetechnologies.com/voltagesensorprobes
Dose-dependent calcium response to muscarnic 1

(M1) receptor agonists and antagonists. CHO M1 cells
were plated in a poly-D-lysine coated 384-well plate
and incubated overnight. The following day, cells were
assayed for a calcium response to carbachol using
the Fluo-4 Direct Calcium Assay Kit. Cells were
stimulated with agonists, carbachol, MCN-A-343,
bethanechol, oxotremorine, and pilocarpine (A) or cells
were treated with antagonists, scopolamine, telenzipine,
and DAMP to block the calcium response elicited by
114 nM carbachol (EC80 for this receptor) (B). Measurements are given in relative fluorescent units (RFU) as
the maximum response minus the minimum response
divided by the minimum response. Rank order of agonist
and antagonist potency agreed with published results.
FLAsH and ReAsH tetracysteine-based protein detection
The TC-FlAsH II and TC-ReAsH II detection kits (Cat. Nos. T34561, T34562) and Gateway Expression Vectors (Cat. No. T34563)
offer a tag-based fluorescence labeling technology for live cell imaging and more. The FlAsH-EDT2 and ReAsH-EDT2 labeling
reagents become fluorescent when they bind to recombinant proteins containing the tetracysteine (TC) motif Cys-Cys-Pro-GlyCys-Cys. FlAsH and ReAsH technology provides sensitive live-cell imaging and subcellular localization of proteins containing the
TC-tag using fluorescence microscopy.
The TC-FlAsH II and TC-ReAsH II detection kits are ideal for protein localization or
real-time protein production studies, though its versatility offers a range of benefits:
Rapid protein detectioneasily constructed six-amino acid tag

Multiplexing flexibilitychoice of red or green fluorescence from the same
tagged protein
Small tag sizereduces interference with target protein

Stable but non-covalent bindingallows dual labeling, pulse-chase experiments
and other dynamic, real-time studies
Compatible with live- or fixed-cell imaging and multiplexing
Multiple applications beyond direct labeling for imaging
Affinity purification
CHO-K1 cells expressing a tetracysteine-tagged

version of -tubulin, labeled with TC-ReAsH
reagent. Upon treatment with vinblastine, a compound
known to perturb cytoskeletal structure, tubulin
drastically rearranges from a reticular structure (A)
to rod shapes (B).
Learn more at lifetechnologies.com/flashreash.
88
Cell analysis
Probes for visualizing

cell structure
This section highlights our most popular and easiest-to-use fluorescent dyes
and fluorescent protein-based reagents for staining organelles and subcellular
structures. Additional information is available online; go to lifetechnologies.com/
organellestainsselectionguide to find selection guides for over 100 different fluorescent probes, organized by these cellular structures:
Adiposomes
Cytoplasm
Cytoskeleton
Endoplasmic reticulum
Golgi complex
Intracellular membranes
Lysosomes
Membrane trafficking
Mitochondria
Nuclear envelope
Nucleoli
Nucleus
Peroxisomes
Plasma membrane
Whole-cell stains for image segmentation in HCS applications
Information about related products, including organelle-specific antibodies

A link to our interactive Cell Staining Simulation Tool that allows you to select
compatible sets of fluorescent dyes for your experiment
Access to immunofluorescence protocols, the Fluorescence Spectra Viewer, the

Image Gallery, and more
Cell structure
Nucleus
Nucleolus
Golgi complex
Tubulin
Endoplasmic
reticulum
Lysosome
Mitochondria
Cytosol
Peroxisome
Actin
Lipid rafts
Plasma
membrane
89
Cell analysis
Cytoskeleton
The cytoskeleton is an essential component of a cells structure and one of the easiest to label with fluorescent reagents. We
offer labeling reagents for monomeric actin (G-actin), filamentous actin (F-actin), tubulin, and other cytoskeletal proteins.
Read more about the options for detecting actin, intermediate filaments, and tubulin in our BioProbes magazine article at
lifetechnologies.com/actintubulindetection. Find a more detailed version of the selection guide below at lifetechnologies.com/
organellestainsselectionguide.
Product
Popular products (fluorescence color)
Key attributes
Alexa Fluor dyelabeled

phalloidins
A22281 (blue), A12379 (green), A34055

(orange), A12381 (orange), A34054 (far
red), A22287 (far red), A22286 (near IR)
Alexa Fluor dyelabeled phalloidins bind to F-actin in nanomolar

concentrations and show equal affinity for large and small filaments;
phalloidins are typically used to stain fixed cells; live-cell staining
requires liposome delivery or microinjection of conjugates,
compromised membranes, or unlabeled phalloidins; contrast between
stained and unstained areas is extremely large; phalloidin-labeled
actin remains functional.
Learn more about phalloidin conjugates for staining actin at
lifetechnologies.com/phalloidinconjugates.
Rhodamine phalloidin
R415 (orange)
Conjugated to a popular orange-fluorescent dye,

tetramethylrhodamine (TRITC); convenient probe for labeling,
identifying, and quantitating F-actin in formaldehyde-fixed and
permeabilized tissue sections, cell cultures, or cell-free experiments;
rhodamine phalloidin will stain live cells.
CellLight Actin GFP
C10582 (green)
Fusions of fluorescent proteins to the N-terminus of -actin in

mammalian cells; allow visualization of cytoskeletal components in
live cells or cytoskeletal rearrangements in HCS cytotoxicity assays.
Learn more about CellLight reagents at
lifetechnologies.com/celllightlabelingproducts.
CellLight Tubulin-GFP,
CellLight Tubulin-RFP
C10613 (green), C10614 (red)
Fusions of fluorescent proteins to the N-terminus of -tubulin that

allow live-cell imaging of microtubules; to monitor tubulin changes as
an indicator of cytotoxicity; also for following cell division in transiently
transduced cells, including primary and stem cells.
Cell structure
CellLight MAP4-GFP,
CellLight MAP4-RFP
C10598 (green), C10599 (red)
Fusions of fluorescent proteins to microtubule associated protein

4 (MAP4), enabling visualization of microtubules in live cells that
can also readily be fixed for endpoint studies such as HCS-based
cytotoxicity analyses.
TubulinTracker Green
T34075 (green)
Live bovine pulmonary arterial endothelial cells were

labeled with CellMask Deep Red plasma membrane
stain (Cat. No. C10046), the tubulin-selective TubulinTracker Green dye (Oregon Green 488 Taxol,
bis-acetate, Cat. No. T34075), mitochondrion-selective
MitoTracker Red CMXRos dye (Cat. No. M7512), and bluefluorescent Hoechst 33342 nuclear stain (Cat. No. H21492).
90
Results in green-fluorescent staining of polymerized tubulin in live cells.
Muntjac skin fibroblasts stained with Alexa Fluor

488 phalloidin, (Cat. No. A12379) an antia-tubulin
antibody (Cat. No. A11126) pre-labeled with Zenon
Alexa Fluor 568 Mouse IgG1 Labeling Kit (Cat. No.
Z25006) and the anticdc6 antibody pre-labeled with
Zenon Alexa Fluor 350 Mouse IgG1 Labeling Kit
(Cat. No. Z25000).
The actin cytoskeleton of the Muntjac fibroblast was

visualized using the green-fluorescent Alexa Fluor
488 phalloidin (Cat. No. A12379). The nucleus was
stained with TO-PRO-3 reagent (Cat. No. T3605).
Cell analysis
Mitochondria
Mitochondria are found in eukaryotic cells, where they make up as much as 10% of the cell volume. They are pleomorphic organelles with structural variations depending on cell type, cell-cycle stage, and intracellular metabolic state. The key function of
mitochondria is energy production through oxidative phosphorylation and lipid oxidation. We offer these probes for live-cell staining
of mitochondria. Find out more at about these products and about anti-OxPhos complex antibodies for fixed cell staining at
lifetechnologies.com/organellestainsselectionguide.
Key attributes
MitoTracker Green FM
M7514 (green)
Live-cell stain; staining does not appear driven by mitochondrial

function. Useful counterstain for mitochondrial mass. Has been
reported to exhibit specific staining after fixation in certain cells.
MitoTracker Red FM,

MitoTracker Orange
CMTMRos, MitoTracker
Red CMXRos, MitoTracker
Deep Red FM
M7510 (orange), M22425 (red), M7512

(red), M22426 (deep red)
Good live-cell stains, and well suited for multicolor labeling

experiments; all are well retained after fixation except MitoTracker
Red FM.
MitoTracker Orange
CM-H2TMRos,
MitoTracker Red
CM-H2XRos
M7511 (orange), M7513 (red)
Do not fluoresce until they enter live cells, where they are oxidized
to the fluorescent mitochondrion-selective probe MitoTracker Red
CM-H and then sequestered in mitochondria.
CellLight MitochondriaGFP,
CellLight MitochondriaRFP
C10508 (green), C10505 (orange)
Fluorescent proteins targeted specifically to mitochondria via the

leader sequence of E1 pyruvate dehydrogenase; delivered to live cells
using BacMam delivery technology.

JC-1
T3168 (green as monomer, red as

aggregate)
Widely used cationic dye that is dependent upon mitochondrial

membrane potential for accumulation; JC-1 monomer (low
concentration) is green, whereas accumulation at higher concentration
shifts emission to the red, allowing ratiometric membrane potential
measurements.
Image-iT LIVE Mitochondrial and Nuclear Labeling

Kit
I34154 (contains two stains: blue and

red fluorescent)
Contains MitoTracker Red CMXRos dye and Hoechst 33342 for highly
selective mitochondrial and nuclear staining in live GFP-transfected
cells.
MitoSOX Red Indicator
M36008 (red)
This indicator is a cationic derivative of dihydroethidum and has

been designed for highly selective detection of superoxide in the
mitochondria of live cells.
Bovine pulmonary artery endothelial cell (BPAEC).

MitoTracker Red CMXRos (Cat. No. M7512), SYTOX
Green nucleic acid stain (Cat. No. S7020), biotin-XX
goat antimouse IgG antibody and Cascade Blue
NeutrAvidin biotin-binding protein (Cat. No. A2663).
Bovine pulmonary artery endothelial cell (BPAEC).

Oregon Green 514 goat antimouse IgG antibody,
(Cat. No. O6383) MitoTracker Red CMXRos (Cat. No.
M7512) and DAPI (Cat. No. D1306).
Cell structure
Product
HeLa cell labeled with CellLight Mitochondria-GFP

(Cat. No. C10600) and CellLight Talin-RFP (Cat. No.
C10612) reagents and with Hoechst 33342 nucleic acid
stain (Cat. No. H21492).
91
Cell analysis
Lysosomes and peroxisomes
Lysosomes are single membrane-bound vesicles that contain digestive enzymes such as glycosidases, acid phosphatases, elastase, cathepsins, carboxypeptidases and a variety of other proteases. The function of this organelle is to break down waste products and cellular debris. Peroxisomes are membrane-bound organelles in the cytoplasm of the cell. They are responsible for
removing harmful peroxides from the cell and also play a role in fatty acid metabolism. We offer these probes for live-cell staining
of lysosomes and for live- and fixed-cell staining of peroxisomes. Find out more about these products and about antibodies to lysosome and peroxisome proteins for fixed-cell staining at lifetechnologies.com/organellestainsselectionguide.
Product
Key attributes
LysoTracker Blue DND-22
L7525 (blue)
Highly selective for acidic organelles at nanomolar concentrations,

shows minimal staining of mitochondria, for labeling and tracking of
acidic organelles in live cells, loads rapidly.
LysoTracker Green
DND-26
L7526 (green)
Is taken up very rapidly by live cells; imaging using this dye is best
between 1 and 5 minutes after dye is added.
LysoTracker Red DND-99
L7528 (red)
Rapid loading, highly selective for acidic organelles at nanomolar

concentrations, show minimal staining of mitochondria, for labeling
and tracking of acidic organelles in live cells.
LysoSensor Blue
DND-167, LysoSensor
Green DND-153
L7533 (blue), L7534 (green)
LysoSensor dyes are live-cell stains that are nonfluorescent at pH 7

and exhibit a pH-dependent increase in fluorescence upon acidification.
Image-iT LIVE Lysosomal

and Nuclear Labeling Kit
I34202 (contains two stains: blue and

red fluorescent)
Contains LysoTracker Red DND-99 dye and Hoechst 33342 for highly
selective staining of lysosomes and nucleus in live GFP-transfected
cells.
CellLight Lysosomes-GFP, C10596 (green), C10597 (red)

CellLight Lysosomes-RFP
Fluorescent proteins targeted to lysosomal associated membrane

protein 1; for live-cell visualization of lysosomes and any process in
which they are involvedphagocytosis, endocytosis, autophagy.
Learn more about CellLight reagents at lifetechnologies.com/
celllightlabelingproducts.
CellLight Peroxisome-GFP C10604 (green)
Fluorescent protein targeted specifically to the peroxisome via the

peroxisomal C-terminal targeting sequence; for live-cell staining.
Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts.
Cell structure
SelectFX Alexa Fluor 488

Peroxisome Labeling Kit
S34201 (green)
Live bovine pulmonary artery endothelial cells (BPAEC) stained with LysoTracker Red DND-99 (Cat. No. L7528). Then, a solution of green fluorescent
dihydrorhodamine 123 (Cat. No. D632, D23806) and blue-fluorescent Hoechst
33258 (Cat.No. H21491) was added and allowed to incubate with the cells for an
additional 10 minutes before the cells were subsequently washed and visualized.
92
Kit for peroxisome labeling in fixed cells, providing a primary antibody

directed against peroxisomal membrane protein 70 (PMP 70), an
Alexa Fluor 488labeled secondary antibody, PBS, blocking solution,
fixative, and permeabilization solutions.
Imaging autophagy in live HeLa cells with CellLight reagents for mitochondria
and lysosomes. Cells were transduced with CellLight Lysosomes-GFP (Cat. No.
C10596) and CellLight Mitochondria-RFP (Cat. No. C10601). Following treatment
with chloroquine to induce autophagosome accumulation, nuclei were stained with
Hoechst 33342 (Cat. No. H21492) and live-cell imaging was performed.
Cell analysis
Endoplasmic reticulum and Golgi apparatus
The endoplasmic reticulum (ER) and Golgi apparatus are primarily responsible for the proper sorting of lipids and proteins in cells.
Consequently, most of the cell-permeant probes for these organelles are either lipids or chemicals that affect protein movement.
The flattened membranous sacs of the ER and the Golgi apparatus can be stained with a variety of lipophilic probes and then
distinguished by their morphology. Find out more about these products and about antibodies to endoplasmic reticulum and Golgi
apparatus proteins for fixed-cell staining at lifetechnologies.com/organellestainsselectionguide.
Product
Key attributes
ER-Tracker Green,
ER-Tracker Red
E34251 (green), E34250 (red)
Live-cell permeant and highly endoplasmic reticulum-selective

BODIPY dyes; when cells are stained using the provided protocol,
staining patterns are partially retained after aldehyde fixation.
CellLight ER-GFP,
CellLight ER-RFP
Fluorescent proteins targeted specifically to the nucleus via the endoplasmic reticulum organelle signal sequence of calreticulin and KDEL
(ER retention signal).
S34200 (green)
Kit for endoplasmic reticulum organelle labeling in fixed cells,

providing a primary antibody directed against ER-associated disulfide
isomerase (PDI), an Alexa Fluor 488 dye-labeled secondary antibody,
PBS, blocking solution, fixative and permeabilization solutions.
Concanavalin A Alexa
Fluor 594 conjugate
C11253 (red)
Fixed cell stain for ER staining; concanavalin A selectively binds to

-mannopyranosyl and -glucopyranosyl residues.
BODIPY FL C5-ceramide,
complexed to BSA
D3521, B22650 (green)
Selective staining of the Golgi complex with applications for lipid

metabolism and trafficking studies; brighter, more photostable than
NBD conjugates; concentration-dependent shift from green to red in
the Golgi complex; good for outlining cell boundaries for observation of
morphogenetic movements and for measuring rates of lipid synthesis
by Schwann cells.
BODIPY TR ceramide,
complexed to BSA
D7540, B34400 (red)
Selective staining of the Golgi complex with applications for lipid trafficking studies, brighter and more photostable than the NBD conjugates; displays no concentration-dependent emission.
Lectin HPA Alexa Fluor

488 conjugate
L11271 (green), L32454 (red)
Lectins selectively bind to terminal -N-acetylgalactosaminyl

residuesan intermediate sugar added in O-linkage to serine and
threonine residues in cis-Golgi cisternae and then substituted with
galactose and sialic acid in the trans-Golgi organelle; for each cell type
or tissue it is necessary to empirically determine appropriate lectins
and staining conditions.
CellLight Golgi-GFP,
CellLight Golgi-RFP
Fluorescent proteins targeted specifically to the Golgi complex via the

human Golgi-resident enzyme N-acetylgalactosaminyltransferase-2.
Cell structure
SelectFX Alexa Fluor 488

ER Labeling Kit
Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts.

NBD C6-ceramide
N1154 (green)
Organelle staining of live bovine pulmonary artery

endothelial cells. Endoplasmic reticulum was labeled
with ER-Tracker Green (Cat. No. E34251); mitochondria were visualized with MitoTracker Red CMXRos
(Cat. No. M7512).
Classic structural marker for the Golgi complex.
U2OS osteosarcoma cells labeled with CellLight

ER-GFP reagent (Cat. No. C10590) and with Hoechst
33342 nucleic acid stain (Cat. No. H21492).
93
Cell analysis
Nucleus
The nucleus of the cell is responsible for regulation of gene expression and is a membrane bound organelle that is comprised of
the nuclear envelope, nucleoli, and nuclear lamina. Find out more about these products and about antibodies to nuclear proteins
for fixed cell staining at lifetechnologies.com/organellestainsselectionguide.
Product
Key attributes
NucBlue Live Cell Stain

(Hoechst 33342 Special
Formulation)
R37605 (blue)
Ready-to-use liquid DAPI formulation in a dropper bottle; excellent cell

permeable nuclear counterstain for live cells.
NucBlue Fixed Cell Stain

(DAPI Special Formulation)
R37606 (blue)
Ready-to-use liquid DAPI formulation in a dropper bottle; excellent

nuclear counterstain for fixed cells.
7-AAD
A1310 (red)
Nuclear stain, GC-selective, weakly permeant in some cell types.
Ethidium homodimer-1
E1169 (red)
One of the highest-affinity fluorescent probes available for nucleic acid

staining; binds to dsDNA, ssDNA, RNA, and oligonucleotides with a
significant fluorescence enhancement (>40-fold); the dyes intercalation is not sequence-selective.
Propidium iodide
P3566 (red)
One of the most commonly used fixed-cell nuclear stains.
Organelle Lights
Nuc-GFP, CellLight
Nucleus-CFP, CellLight
Nucleus-GFP, CellLight
Nucleus-RFP
O36209 (green), C10616 (blue), C10602

(green), C10603 (red)
Fluorescent proteins targeted specifically to nucleus via the SV40

nuclear localization sequence; allows visualization of the nucleus
without utilizing DNA-binding dyes; can also be used as a segmentation marker in endpoint HCS studies.
CellLight Histone 2B-GFP, C10594 (green), C10595 (red)

CellLight Histone 2B-RFP
Alternatives to DNA staining for localizing the nucleus and chromosomes; allow localization of histones and following of cell division in
live cells.
Cell structure

SelectFX Nuclear Labeling S33025 (several)
Kit, for fixed cells (DAPI,
SYTOX Green, 7-AAD,
TO-PRO-3 iodide)
A sampler kit of our most popular nuclear stains for fixed cells, with
optimized protocols for use.
SYTO 9, SYTO 14, SYTO

82, SYTO 59, SYTO 61
S34854 (green), S7576 (green), S11363

(orange), S11341 (red), S11343 (red)
Live-cell nuclear counterstains; permeant to eukaryotic and prokaryotic cell membranes; SYTO 9 is useful to indicate bacterial viability;
SYTO 61 stains live and dead bacteria; SYTO 14 is twice as fluorescent when complexed with RNA as with DNA.
SYTOX Green nucleic acid

stain
S7020 (green)
Highest-affinity SYTOX stain, fixed-cell stain, useful as a dead-cell

stain, 500x increase in fluorescence upon nucleic acid binding, can be
used in conjunction with blue and red stains for multiplexing.
SYTOX Orange nucleic acid S11368 (orange)

stain
Fixed-cell nuclear stain, some mitochondrial staining, shorter-wavelength emission than propidium iodide, useful as a dead-cell stain,
500x increase in fluorescence upon nucleic acid binding, can be used
in conjunction with blue and green stains for multiplexing.
TO-PRO-3 iodide
Ultrasensitive dsDNA detection, dead-cell stain, electrophoresis

prestain, used as an alternative to annexin conjugates for analyzing
apoptotic cells.
T3605 (far red)
U2OS osteosarcoma cells transduced with CellLight Histone 2B-RFP (Cat. No. C10595) and CellLight MAP4-GFP (Cat. No. C10598) reagents.
94
Cell analysis
Probes for cell tracking

and neuronal tracing
Molecular Probes fluorescent cell trackers and neuronal tracers are essential tools for following cells in complex environments, and
have been used extensively in studies of cell migration, wound healing, and stem cell differentiation. These probes (Table 1) are stable,
nontoxic at working concentrations, well retained in cells (from hours to days), and brightly fluorescent at physiological pH. Moreover,
they are available in fluorescence emission colors that span the spectrum, allowing simultaneous detection of cell movement and other
critical cell functions. Learn more about all of our fluorescent cell trackers and neuronal tracers at lifetechnologies.com/celltracing.
Table 1. Molecular Probes fluorescent cell tracking probes and tracers.
Probe class
Retention time
Fixable General attributes
Instrument
platform * Selected Cat. Nos.
Calcein AM
probes
<1 day (13hr)
No
Acetoxymethyl (AM) estermodified compounds that can passively

diffuse across cell membranes, where nonspecific esterases cleave
the AM ester moieties to produce a product that is retained only in
live cells.
FC, FM
For imaging: C3099,

C3100MP, C1430, C34852,
For flow cytometry:
C34858, C34853, C34851
CellLight
reagents
Days to weeks or Yes

68 generations
Baculovirus-packaged DNA constructs for the expression of

nontargeted fluorescent proteins or of fluorescent proteins fused to
signal peptides for specific targeting to key cytoskeletal structures
and organelles.
FC, FM,
HCS
B10383, C10582, C10601
Cell-permeant, thiol-reactive labeling reagents containing a

chloromethyl moiety that can react with intracellular thiol groups from
cysteine residues in proteins or glutathione.
FC, FM,
HCS
C7025, C2925, C34552,

C2927
CellTracker 68 days or
probes
68 generations
Yes
510 days or
Yes
810 generations
Cell-permeant, amine-reactive labeling reagents containing a

succinimidyl ester that forms stable adducts with primary amines
located intra- and extracellularly.
FC, FM
C34557, C34554, C34553
Qtracker
Cell Labeling
Kits
Days to weeks or Yes

810 generations
Provides Qdot probes that contain a selective targeting peptide for

uptake into the cytoplasm of live cultured cells. Once inside cells, Qdot
probes provide intense, stable fluorescence that can be traced through
several generations.
FC, FM,
HCS, M
Q25021MP, Q25041MP,
Q25071MP, Q25001MP,
Polar tracers
Hours to days
Some
Highly water-soluble dyes and other detectable probes that can be

used as cell tracers. In most cases, the polarity of these probes is too
high to permit them to passively diffuse through cell membranes.
Consequently, special methods for loading the dyes into cells must be
employed, including microinjection, pinocytosis, or techniques that
temporarily permeabilize the cell membrane.
FM
A12924, A12923, A12922,

A10436, A20501MP,
A10436, A20502, B1592
Membrane
tracers
Hours to weeks
Some
FM, FC
Lipophilic dialkylcarbocyanines, including DiI, DiO, DiD, and DiR, are
widely used retrograde and anterograde tracers in live and fixed tissues.
Ready-to-use NeuroTrace DiI and DiO tissue-labeling pastes can be
applied directly to tissues using the tip of a needle, allowing dye penetration into bundled neurons for labeling axons both on and below the
surface. Also available are the NeuroTrace Multicolor Tissue-Labeling
Kit, which contains DiI, DiO, and DiD pastes, and the Lipophilic Tracer
Sampler Kit, which contains DiI, DiO, DiD, DiR, and DiA analogs.
D282, D3899, D3911,

D7756, N22880, D275,
D3898, N22881, D307,
D7757, D12731
Fluorescent
dextran
conjugates
Hours to weeks
Yes
Dextrans are hydrophilic polysaccharides characterized by their

moderate to high molecular weight, good water solubility, and low
toxicity.
FM, FC
D22910, D1956, D3308,

D1830, D22914, D7139,
D22913
Fluorescent
Hours to days
microspheres
Some
Polystyrene fluorescent microspheres that can be used for a wide

range of applications including blood flow determination, tracing, in vivo
imaging, and calibration of imaging and flow cytometry instruments.
FM, FC
F8760, F8770, F8787,

F21012, F20892, F13083
Proteins
& protein
conjugates
Hours to weeks
Yes
Cholera toxin B conjugate is a powerful tool for retrograde labeling of

neurons. Other protein conjugates, such as albumin and ovalbumin,
are used to target receptors, detect protease activity, and measure
intracellular pH.
FM, FC,
HCS
C22841, C34775, C22842,

C22843, C34776, C34778,
A13100, A23015
NeuroTrace
Nissl stains
Days to weeks
No
Nissl staining is a standard histochemical method for visualizing

neurons in the brain and spinal cord.
FM
N21479, N21480, N21481,

N21482, N21483
Cell tracing
CellTrace
probes
* FC = flow cytometry. FM = fluorescence microscopy. HCS = high-content screening and analysis. M = microplate assay. After fixation, these emGFP and tagRFP fusions can be detected
withanti-GFP antibodiesandanti-RFP antibodies, respectively. Lysine-fixable dextrans contain lysines and can therefore be fixed in place with formaldehyde or glutaraldehyde. Fixable
dextrans contain free amines (but not lysines) and can be fixed in place with formaldehyde or glutaraldehyde.
95
Cell analysis
Calcein AM probes
The traditional nonfixable calcein probes are useful tracers for

both cell tracing and viability studies. These tracers contain
acetoxymethyl (AM) moieties that block the inherent negative
charges on the molecules, allowing their passage across cell
membranes. Once inside the cell, nonspecific esterases cleave
the AM esters to produce cell-impermeant polar molecules that
can be efficiently retained in the cell for several hours. Unlike
the CellTracker and CellTrace probes described below, the
Calcein AM probes are retained only in live cells. When paired
with a cell-impermeant nucleic acid stain such as SYTOX
Green dye or propidium iodide, these calcein tracers are effective for differentiating viable and dead cells using microscopy
(see figure, right) or flow cytometry. Calcein probes are easy to
load and are effective for 2-8 hours, depending on the cell type.
Calcein AM. Jurkat cells were incubated with either green-fluorescent calcein
AM (C1430, C3099, C3100MP), CellTrace calcein red-orange AM (C34851), or
calcein blue AM (C1429). The labeled cells were then combined and imaged with
the appropriate filters.
Control
24 hr post-wound
24 hr post-wound with
0.1 m cytochalasin D
CellTracker probes
Cell tracing
CellTracker fluorescent probes (see figure, right) freely pass

through cell membranes and are converted to cell-impermeant
reaction products. The cell-impermeant form is passed to
daughter cells through several generations but is not transferred to adjacent cells in the population. Cells that are loaded
with CellTracker probes are typically fluorescent and viable
for at least 24 hours, making these probes excellent long-term
cell tracers. Furthermore, the staining pattern can be fixed with
formaldehyde or glutaraldehyde to allow long-term storage of
the labeled cells or tissue, to facilitate safe handling of potentially pathogenic samples, and to permit signal amplification in
cases where the anti-dye antibody is available. CellTracker
probes are not transferred to adjacent cells in a population, and
are ideal long-term tracers for transplanted cells or tissues. The
signal from CellTracker probes is typically monitored using
fluorescence microscopy, and the probes are offered in several
fluorescent colors for compatibility with multicolor analyses.
CellTrace probes
The CellTrace probes are also well retained in cells and are
retained after formaldehyde-based fixation once they have
reacted with proteins or other amine-containing biomolecules.
These amine-reactive cell tracersincluding CellTrace
Violet, CellTrace CFSE, and CellTrace Far-Red DDAO-SE
probeseach contain a succinimidyl ester that reacts with
available primary amines located both inside and outside the
cell. The bright, homogeneous staining of CellTrace Violet
probe shows very little fluorescence variation between cells in
a population, allowing visualization of distinct generations of
proliferating cells in a fluorescence histogram.
Qtracker probes
The Qtracker Cell Labeling Kitsavailable with Qdot nanocrystals in seven brilliant colorscontain the reagents needed
to deliver fluorescent Qdot nanocrystals into live cells,
providing a powerful method for real-time tracking studies over
96
Wound-healing assay with CellTracker Green CMFDA. (Left) After growing in

culture for 72 hr, a monolayer of Gibco human neonatal dermal fibroblasts was
stained with CellTracker Green CMFDA. (Middle) The stained cell monolayer was
scratched and left to heal for 24 hr. Cells have reinvaded the wound but are not as
dense as in the control plate. (Right) The stained cell monolayer was scratched and
then treated with 0.1 M cytochalasin D for 24 hr to disrupt actin polymerization,
resulting in no closure of the wound site.
Qtracker probes. (A) Qtracker 625. (B) Three-color Qtracker cell labeling of 3T3
(green), HeLa (red), and U188 (white) cells labeled with Qtracker 565, 655, and 705,
respectively. (C) Vascular labeling with Qtracker vascular labels.
extended time periods. These Qdot nanocrystals contain a

targeting peptide that allows their selective uptake into the cell,
where they are distributed in vesicles throughout the cytoplasm
(see figure, above).
Inside the cell, the Qdot nanocrystals exhibit an intense, photostable fluorescence that can be observed using continuous
illumination, without time constraints due to photobleaching
or degradation. Their fluorescence is maintained in complex
cellular environments and under various biological conditions,
including changes in intracellular pH, temperature, and metabolic activity, and they are passed to daughter cells through at
least 7 generations, or about 8 days in U2OS cells. Qtracker
Cell Labeling Kits have been used for diverse applications
including in vivo tracking of hematopoietic stem cells, following
Cell analysis
Schwann cells after injection into nerve grafts, and detecting
neutrophil survival in infected skin wounds.
For in vivo vascular imaging in small animals, Qtracker nontargeted quantum dots are designed to be injected into the tail
vein of mice for the study of vascular structure using small
animal in vivo imaging (SAIVI) techniques (see figure). These
nanocrystals exhibit intense fluorescence with red-shifted emission for increased tissue penetration and avoidance of background autofluorescence, and have a PEG surface coating developed to minimize nonspecific interactions and reduce immune
response. Because the PEG surface coating does not contain
reactive functional groups, the Qtracker non-targeted quantum
dots are retained in circulation longer and can be imaged for up
to 3 hours with a single injection, or for longer periods of time
with additional injections.
CellLight reagents
CellLight reagents are ready-to-use fusion constructs of

emGFP and TagRFP for the expression of untargeted, cytoskeleton-targeted, or organelle-targeted fluorescent proteins.
The CellLight cytoplasmic markeralso called BacMam
GFP transduction controlproduces emGFP expression in the
cytosol and nucleus of a wide range of cell types. The transiently
transduced cells typically express the fluorescent protein fusion
for about 5 days in standard cell lines (e.g., HeLa and CHO). With
slowly dividing cellssuch as some primary cellsexpression
has been demonstrated for up to 2 weeks, and with terminally
differentiated neurons we have observed expression for more
than 3 weeks after transduction.
tracers, which evenly distribute throughout the whole cell, are

good tools for total neuronal cell visualization (see figure, below).
These dyes can also be used to visualize gap junctions.
Membrane tracers
Lipophilic dialkylcarbocyanines, including DiI (see figure, below),
DiO, DiD, DiR, FM 1-43, and FM 4-64, can be introduced into
membranes by direct application or with kits (Vybrant celllabeling solutions). Lateral diffusion of these dyes eventually
stains the entire cell. These probes are widely used as retrograde
and anterograde tracers in live and fixed tissues, as well as for
long-term assays of cellcell association, and are available in
a wide selection of fluorescence emission colors and physical
forms (solid, dissolved in organic solvents, suspended in paste,
and large crystals).
Ready-to-use NeuroTrace DiI and DiO tissue-labeling pastes
can be applied directly to tissues using the tip of a needle, allowing
dye penetration into bundled neurons for labeling axons both
on and below the surface. Also available are the NeuroTrace
Multicolor Tissue-Labeling Kit, which contains DiI, DiO, and DiD
pastes, and the Lipophilic Tracer Sampler Kit, which contains DiI,
DiO, DiD, DiR, and DiA analogs.

Cell tracing
Neuronal tracing
Fixable polar tracers
We prepare a wide variety of highly water-soluble dyes and

other detectable probes that can be used as cell tracers. Molecular Probes fixable polar tracers include the membraneimpermeant probes lucifer yellow and Alexa Fluor hydrazides. In most cases, the polarity of these water-soluble probes
is too high to permit them to passively diffuse through cell
membranes. Consequently, special methods for loading the
dyes into cells must be employed, including microinjection,
pinocytosis or techniques that temporarily permeabilize the
cell membrane. These probes are often microinjected, and are
useful for a variety of applications including neuronal tracing,
investigating cellcell and cellliposome fusion, membrane
permeability, transport through gap junctions, and cell uptake
during pinocytosis. These fluorescent hydrazide derivatives are
covalently linked to surrounding biomolecules during aldehyde
fixation, allowing subsequent antibody-based analyses.
Lucifer yellow and sulforhodamine 101 (Orange/Red) are widely
used tracers that must be loaded by injection. These polar
Cultured left-upper quadrant neurons from the sea slug Aplysia californica that
have been microinjected with either lucifer yellow CH (L453, L682, L1177, L12926)
or sulforhodamine 101 (S359). Image contributed by David Kleinfeld, AT&T Bell
Laboratories, and Brian Salzberg, University of Pennsylvania School of Medicine.
DiI, the classic red membrane stain. DiI is nonfluorescent in aqueous solutions,
but has incredibly intense fluorescence in membranes. In this image, DiI (D282,
D3911) is used as a diagnostic tool to evaluate patterns of innervation in a newborn
mouse cochlea. The larger image is of a mutant cochlea and the inset is of a wildtype cochlea. Images contributed by Bernd Fritzsch, Creighton University, and L.
Reichardt and I. Farinas, Howard Hughes Medical Institute, San Francisco.
97
Cell analysis
Fluorescent dextran conjugates
Stains and kits for neuronal tracing
Fluorescent dextran conjugates (such as Alexa Fluor dextrans)

are ideal cell-lineage tracers because they are relatively inert,
exhibit low toxicity, and are retained in cells for long periods.
Dextrans are offered in a range of molecular weights and are
useful size-exclusion probes for determining pore sizes in
membranes. They are typically loaded into cells by microinjection, whole-cell patch clamping, or electroporation.
Fluorescent and biotinylated dextrans are routinely employed

to trace neuronal projections. Dextrans can function efficiently
as anterograde or retrograde tracers, depending on the study
method and tissue type used. Active transport of dextrans
occurs only in live, not fixed tissue. Dextran conjugates with
molecular weights up to 70,000 daltons have been employed as
neuronal tracers in a wide variety of species.
Fluorescent microspheres
Cell tracing
FluoSpheres and TransFluoSpheres polystyrene microspheres are intensely fluorescent, durable, and inert tracers.
These microspheres are particularly useful as long-term
markers for transplantation studies. They are either injected
into cells or taken up by phagocytosis. These microspheres
satisfy several prerequisites of ideal long-term biological
tracers. Because the dyes in our microspheres are incorporated
throughout the microsphere rather than just on its surface, the
fluorescence output per microsphere is significantly greater
than that obtained from protein or dextran conjugates and is
relatively immune to photobleaching and other environmentdependent effects. FluoSpheres and TransFluoSpheres
microspheres are also biologically inert and physically durable,
and they are available with a large number of uniform sizes
and surface properties. Furthermore, their spectral properties
can be freely manipulated during manufacture without altering
their surface properties.
Nissl staining is a standard histological method for retrograde

tracing of neurons in the brain and spinal cord. Composed of
ribosomal RNA associated with the rough endoplasmic reticulum in neuronal perikarya and dendrites, the Nissl substance
redistributes within the cell body in injured or regenerating
neurons, providing a marker for the physiological state of the
neuron. Our NeuroTrace Nissl stains are selective for the
Nissl substance characteristic of neurons, and provides more
sensitivity than traditional histological dyes like toluidine blue
or cresyl violet. These Nissl stains are available in a wide spectrum of fluorescent colors for staining neurons, either alone or
in combination with immunofluorescence staining of specific
proteins.
The BrainStain Imaging Kit (see figure, below) enables 3-color
combinatorial labeling of myelin, neurons, and nuclei in brain
cryosections in a single 20-minute staining step plus washes.
This kit contains novel stains that can be used separately or
together in one staining solution, replacing traditional methods
that can take 13 days. Standard histochemical methods such
as immunohistochemistry are compatible with these stains.
Each BrainStain kit provides:
FluoroMyelin Green fluorescent myelin stain

NeuroTrace 530/615 red-fluorescent Nissl stain
DAPI nuclear counterstain
For more details about Molecular Probes fluorescent cell

tracers, visit lifetechnologies.com/celltracing.
Proteins and protein conjugates

Protein tracers have molecular weights between ~12,000
(cholera toxin subunit B conjugates) and ~240,000 (B- and
R-phycoerythrin). Molecular Probes cholera toxin conjugates
are made from a recombinant version of the B subunit only.
This allows us to provide a high-purity product completely free
of the toxic A subunit. Cholera toxin B subunit (CT-B) attaches
to cells by binding to ganglioside GM1, making it a powerful tool
for retrograde labeling of neurons. This tracer has been used in
a variety of applications, including tracing of rat forebrain afferents, projections of the parabrachial region, and neurons of the
urinary bladder wall. When used in neuronal tracing applications, CT-B is typically introduced by pressure injection or by
iontophoretic injection into neural tissue.
Other protein conjugates, such as albumin and ovalbumin, are
used to target receptors, detect protease activity, and measure
intracellular pH.
98
BrainStain Imaging Kit. Mouse brain cryosections of the hippocampal region

(top) and diencephalon region (bottom), labeled simultaneously with three dyes:
DAPI (nuclei, blue), FluoroMyelin Green stain (myelin, green), and NeuroTrace
530/615 red Nissl stain (neuronal cell bodies, red).
Cell analysis
Tali Image-Based Cytometer

Visualize, quantify, and analyze your cells
Versatilegenerate visual and analytical data on GFP/RFP expression, apoptosis,

and cell viability
Convenientset up and analyze cells in minutes right at your bench, with minimal
cleaning and maintenance
Accurateobtain statistically significant 3-parameter population analysis
The Tali Image-Based Cytometer is a 3-channel benchtop imaging platform that
performs quantitative analysis of GFP/RFP expression, apoptosis, cell viability, and
much moreright at your bench. And, its fast, delivering a typical cell population
analysis in about 45 seconds using only 25 L of sample. The Tali cytometer contains
bright-field, red fluorescence, and green fluorescence channels, enabling researchers
to simultaneously count green- and/or red-fluorescent stained cells, as well as cells
expressing GFP and RFP. The Tali cytometer displays fluorescence data for the
population in histograms, which can be adjusted (gated) to collect the fluorescence of
interest. Both the raw cell-by-cell data and simple reports can then be exported for
archiving and sample comparisons.
The Tali Image-Based Cytometer
The Tali Image-Based Cytometer is ideal for:
Rapid analysis of GFP/RFP vector transfection efficiency

Identification of GFP- and/or RFP-positive cells (see figure below)
Simple two-color apoptosis assays using annexin V and PI (see figure below)
Accurate population analysis of cell viability
Quick verification of samples before sorting or flow cytometry analysis
Find the Tali Image-Based Cytometer, consumables, reagents, accessories, and
application data at lifetechnologies.com/tali.
Instruments and accessories
Simultaneous analysis of GFP- and RFP-expressing cells. The Tali Image-Based

Cytometer displays green and red fluorescence channels, enabling simultaneous
quantitation of green and red fluorescence from proteins and stains. The Tali
cytometer also provides actual cell counts and automatically calculates cell
concentration for each cell population.
The Tali Apoptosis Assay. The Tali Apoptosis Assay uses an annexin VAlexa Fluor
488 conjugate and propidium iodide to assess cell viability and health. Results indicate whether each cell is live (unstained), dead (yellow and red), or apoptotic (green).
99
Cell analysis

The real answer at the speed of sound
Better data in less time with less effort:
Faster acquisitionmore events/second and faster run time/sample

Better sensitivity and precisionlow CVs even at high collection rates
Simplified sample preparationno-lyse, no-wash method eliminates cell loss
and improves data accuracy
Easy to usefree and intuitive software allows you to analyze data from
anywhere at any time
Low cost of ownershiplow fluid consumption and direct count without

counting beads
The Attune Acoustic Focusing Cytometer, the worlds first and only acoustic
focusing flow cytometer, uses ultrasonic waves to align cells into a single focused
line. With the Attune Acoustic Focusing Cytometer, you get both high sample input
rate and high precision. You control the sample concentration, the flow rate, the
number of photons you detect, the length of your experiment, and more, which
allows you to maximize the data from each cell sample. But best of all, the Attune
cytometer makes it easy with streamlined sample prep and easy-to-use acquisition
and analysis software.
Powered by Molecular Probes flow reagents, Attune cytometer offers two laser
configurations (blue/red or blue/violet), and the Attune Autosampler and enables you
to optimize performance and throughput without sacrificing data quality.
The Attune Acoustic Focusing Cytometer has been used to investigate aspects of
cancer biology, stem cells, oceanography, and microbiology. In fact, cells up to 100 m
in diameter have been run without clogging our 200 m flow cell, and the instrument
also handles particles as small as 1 m. Most standard applications for flow cytometry have been tested on the Attune cytometer, including:
Apoptosis
Rare-event detection
Basic phenotyping (up to 6 colors)
Phagocytosis
Cell cycle analysis
Detection of phosphoproteins
Cell proliferation assays
Detection of intracellular markers
Live/dead cell discrimination
No software licensing fees
Attune sample rate: 1,000 L/min
7
6
5
SSC
Research applications supported
4
3
2
CD34
1
0
101
102
103
CD34+
104
105
Identification of CD34+ cells from peripheral blood.

Peripheral blood from a normal donor was stained and
run on the Attune Acoustic Focusing Cytometer at a
collection rate of 1,000 L/min with a stop gate set at
500,000 total cells. A rare population (0.045%) of CD34+
cells (red box) was identified within the population of
cells with an acquisition time of 4 min 28 sec.
The Attune Cytometric Software offers limited copies and can be downloaded without
licensing fees. You can analyze the results from any computer at your convenience at
any time from anywhere, and an intuitive user interface eases the learning curve.
Simple, user-defined analysis

Automated or user-defined compensation
Automated experimental settings that can be completely customized and saved for
future experiments
Automated instrument checks

Recover unused samples (tube only)
Learn about the advantages of acoustic focusing technology and the Attune
cytometer at lifetechnologies.com/attune.
100
106
Cell analysis
New Molecular Probes reageNts
FLoid
Cell Imaging
Station
siMPlify
fluoresceNt
iMagiNg
Click. Print. Love. Yes, its that easy
Perfect for a quick check of your stained cells, the FLoid Cell Imaging Station is
designed to deliver high-quality, 3-color fluorescent cell images right at your benchtop
with an interface so simple even novice users can collect data in just a few clicks of
the mouse.
Simplified protocols
Ready to use with no dilutions or calculations
Simplecollect images in no time with the intuitive user interface

Practicalprint cell images and place them in your notebook
Accessiblecapture fluorescent cell images at your bench, not in the darkroom
Robustprotect expensive confocal microscopes from excessive day-to-day use
Informativefind the most popular Molecular Probes reagents
Convenient packaging for easy storage
Meet FLoid at lifetechnologies.com/floid.
The FLoid Cell Imaging Station
backDropbackgroundsuppressor
ReadyProbes reagents
Effectively suppresses background

fluorescence for live cell imaging
Focus on your data, not reagent preparation with new ReadyProbes

imaging reagents
No pipetting, calculations, or dilutions
Quickuser-friendly dropper-bottle formulations with minimal

or no dilutions
Intuitiveclear, concise, streamlined protocols

Accessibleroom temperaturestable reagents
Place right on your bench where you need it

Immediate reduction of blue, green or red
background signal
BPAE Molecular
cells labeledProbes
with CellLight
Golgi-RFP inreagents
DMEM, 20% FBS without
Find
ReadyProbes
(A) and with (B) BackDrop Background Suppressor
at lifetechnologies.com/readyprobes.
image-itfixation/PermeabilizationKit
Complete, ready to use kit to fix, permeabilize and wash your

No need to prepare fresh fixative
Room temperature storage
Compatible with fluorescent proteins, dyes, antibodies, and

iT reagents
labeling
BPAE
with and
red-fluorescent
Texasusing
Red-X
BPAE cells
cellsstained
were fixed
permeabilized
thephalloidin
Image-iTfor
F-actin,
mouse monoclonal anti-tubulin
in conjunction with green-fluorescent
FixationPermeabilization
Kit.
BODIPY FL goat antimouse IgG for labeling microtubules, and blue-fluorescent
DAPI for labeling the nuclei. Image was captured using the FLoid Cell Imaging
Station. Prior to immunostaining, cells were fixed and permeabilized with the readyto-use Image-iT Fixation/Permeabilization Kit (Cat. No. R37602).
101
Cell analysis
Primary antibodies for

flow cytometry
Your solution for multicolor flow cytometric analysis

We offer a diverse array of highly specific RUO, ASR, and IVD primary antibodies to targets
studied in both basic and clinical research settings. These antibodies are validated for flow
cytometry and include specificities for CD markers, cytokines, chemokines, and growth
factors, as well as antibodies with utility for oncology, immunology, cell signaling, apoptosis,
proliferation, and stem cell research. From subset identification of heterologous cell
populations to rare-event detection, multicolor flow cytometry can help answer complex cell
biological questionsin less time, with less sample.
Flexiblebroadest range of dye, protein, and Qdot nanocrystal antibody conjugates

Compatiblewide breadth of specificities for numerous types of applications
Reliablequality products for clinical and research applications, RUO, ASR, and IVD
reagents available
Trustedreferred to in over 20,000 journal citations
We offer 24 different fluorescent dyes (including the Alexa Fluor family of dyes, Pacific Blue
and Pacific Orange dyes), proteins (PE, PerCP, and APC), and Qdot nanocrystals conjugated
to a wide range of antibodies, including ten different tandem conjugates to further expand your
multiparametric flow cytometry capabilities.
Go to lifetechnologies.com/flowantibodies to find:
Product details for over 950 antibodies validated for use in flow cytometry, including
human, mouse, rat, and monkey specificities
Our online Flow Cytometry Resource Center (lifetechnologies.com/flowresources) that
Primary antibodies for flow cytometry
allows you to access our video tutorials, webinars, protocols, links to published literature
and more
Our web-based Primary Antibody Selection Tool that allows you to search for antibodies by
protein target, species reactivity, host, label, and application
A downloadable Fluorophores for Flow Cytometry Selection Guide to help you find dyes
that are compatible with 20 different commercially available cytometers
A link to our Flow Cytometry mobile app
102
Cell analysis
Simplify multiplexing with our broad color selection
150
100
50
104
4.5%
102
1044
1055
10
-629
104
0.0%
103
Qdot 605-CD3 (605/20)
Qdot 605-CD4 (605/20)
0.0%
105
103
104
-807
103
104
105
APC-Alexa Fluor 750-CD8 (780/60)
103
105
105
6.9%
104
1.3%
103
103
104
RPE-CD16+CD56 (575/26)
0.0%
103
0
-529
-807
103
104
105
6.7%
105
104
1.5%
103
-807
105
103
104
105
Fluorescein-CD2 (525/50)
105
1.0%
104
59.6%
103
Gated: lymphocyte
+ monocyte
-633
-633
-1,969
0.0%
104
APC-Alexa Fluor 750-CD8 (780/60)
-629
-629
0.9%
Qdot 605-CD4 (605/20)
Qdot 655-CD3 (655/20)
1033
10
Qdot 705-CD45 (710/30)
104
-533
-533
1022
10
105
0.0%
105
Qdot 605-CD4 (605/20)
Side scatter
200
1.7%
105
Qdot 655-CD3 (655/20)
Qdot 655-CD3 (655/20)
250
Qdot 655-CD3 (655/20)
The availability of more color options allows more efficient experiments, gaining more
information with less sample, in less time. Through multiplexing, experimental error is
diminished by minimizing the number of samples used to obtain results. Our wide selection of fluorophores allows researchers to select panels with minimal spectral overlap.
-4,882
-103 0
103
104
RPE-Cy7-CD19 (780/60)
105
-265
0 102
103
104
105
APC-CD14 (670/14)
Human PBLs were labeled with anti-CD4 Qdot 605, anti-CD3 Qdot 655, anti-CD45 Qdot 705, anti-CD2 FITC, anti-CD16+CD56 RPE, anti-CD19 RPE-Cy7, anti-CD14
APC, and anti-CD8 APC-Alexa Fluor 750. Samples were run on an LSR II flow cytometer. Plots are gated on lymphocytes by side scatter and CD45, except as noted.
Axes are labeled with the filters used; plots are labeled with compensation values.
103
Cell analysis
Fluorophores to fit every common laser and instrument configuration
By leveraging years of expertise in the creation and optimization of various technologies including organic dyes, fluorescent
proteins, and Qdot nanocrystals, we are able to offer a complete portfolio of Molecular Probes fluorescent labels that spans the
near-UV, visible, and near-IR spectrum. Regardless of your instrument and its various on-board excitation sources, we have labels
designed to help you get the most out of every sample and every flow cytometry run.
Fluorophore selection guide for flow cytometry.

Laser
Fluorophore
UV
Violet
Alexa Fluor 405
Pacific Blue
Pacific Orange
532 nm
561 nm
Emission (nm)
421
455
551
PerCP
R-phycoerythrin
(R-PE, PE)
575
PE-Texas Red
615
PE-Alexa Fluor 610
628
TRI-COLOR (TC,
PE-Cy5)
670
PE-Cy5.5
694
Qdot 655
Qdot 705
Qdot 800
Alexa Fluor 488
Fluorescein
PE-Alexa Fluor 700

PE-Cy7
800
723
767
Alexa Fluor 700

APC-Alexa Fluor 750
705
678
PE-Alexa Fluor 700
APC-Cy5.5
PerCP-Cy 5.5
655
525
Alexa Fluor 647
605
519
Allophycocyanin (APC)
104
633/635 nm
Qdot 605
488 nm
660
668
694
696
775
695
723
Cell analysis
Qdot nanocrystals for multicolor flow cytometry
Qdot nanocrystals are nanometer-scale semiconductor particles with unique fluorescence properties that
make them attractive labels for multicolor flow cytometry. These fluorophores are optimally excited by a UV
or violet (405407 nm) laser, but can also be excited with longer-wavelength light sources. Qdot nanocrystals are brighter than most organic fluorophores when compared to conventional dyes. And unlike conventional dyes that photobleach over time, Qdot conjugates remain fluorescent under constant illumination.
This fluorescence stability allows ample time for flow cytometric analysis and permits additional analysis
steps after sorting.
With Qdot nanocrystals you get:
Multiplexing capabilityadd Qdot nanocrystal conjugates to existing marker experiments to increase

the number of detectable parameters
StabilityQdot nanocrystals do not degrade over time like tandem conjugates, which promotes better
reproducibility
Flexibilityefficiently excited at 405 nm or 488 nm, maximizing violet laser use

Minimal single-laser compensationnarrow emission spectra allow for minimal compensation when
using a single excitation source
Learn more about Qdot nanocrystals and their applications in flow cytometry and find Qdot nanocrystal conjugates of antibodies that recognize human CD markers at lifetechnologies.com/qdotinflow
and find out more about other fluorescent tools that are compatible with your instruments violet laser at
lifetechnologies.com/violettools.
1. Qdot 525 conjugate excitation
12,000,000
10,000,000

8,000,000

6,000,000
4,000,000
1. Qdot 525 conjugate emission
2,000,000

0
400
500
600
700
Wavelength (nm)
800
900
1,000
Extinction coefficient (M-1cm-1)
2. Qdot 565 conjugate excitaiton

Extinction coefficients and emission profiles for selected Qdot nanocrystals. Excitation is presented as extinction coefficient; emission is
normalized to maximum peak height.
Fixation and permeabilization for flow cytometry samples
By treating sequentially with the two reagents in the FIX & PERM kits, cells undergo mild fixation and
permeabilization that leaves the morphological scatter characteristics intact. This enables researchers to
accurately identify previously undetectable intracellular markers, such as cytoplasmic or nuclear enzymes,
oncoproteins, cytokines, and immunoglobulins.
FIX & PERM reagents:
Are designed for use with all commercially available flow cytometers
Allow the efficient detection of a wide variety of markers and intracellular proteins
Help reduce the amount of background staining observed
Find protocols and ordering information online at lifetechnologies.com/fixperm.
105
Cell analysis
Secondary antibodies and

labeling strategies
Molecular Probes labeling and detection

products
We offer a wide variety of Molecular Probes labeling and detection products to aid
researchers in producing the best images possible. Some of our most popular productsAlexa Fluor dyelabeled secondary antibodiescan be found in labs across the world and have been foundational tools for
fluorescence imaging for decades. We have also developed a number of kits that make it easy for you to directly label your own
antibodies for use in fluorescence microscopy and other applicationsfrom the 10-minute labeling offered by Zenon technologies
to kits that allow you to label anywhere from 10 g up to 1 mg of antibody. We have developed streptavidin conjugates that deliver
moderate signal amplification from your stained samples and Tyramide Signal Amplification (TSA) kits that allow you to detect
very low-abundance targets.
Learn about all of the Molecular Probes options for immunodetection at lifetechnologies.com/antibodies.
Immunodetection strategies
Use the table below to choose the best immunodetection strategy for your imaging experiment.
Strategy
Direct detection with

labeled antibody
Labeled secondary
antibody to detect a bound
(unlabeled) primary
antibody
Streptavidin or other
avidin products to detect
biotinylated primary or
secondary antibody for
moderate amplification
Enzymatic signal
amplification for greater
amplification
Example product
Alexa Fluor 488 Antibody

Labeling Kit
(Cat. No. A20181)
Alexa Fluor 488 Goat

Anti-Mouse IgG (H+L)
(Cat. No. A11001)
Streptavidin, Alexa Fluor

488 conjugate
(Cat. No. S32354)
TSA Kit #22, with HRPstreptavidin and Alexa

Fluor 488 tyramide
(Cat. No. T20932)
Fluorophore-conjugated
secondary antibodies
Biotinylated
secondary antibodies
Enzyme amplification
Antigen
Primary Ab
Directly conjugated
primary antibodies
Secondary Ab
Biotin streptavidin
complex
Enzyme
Fluorophores
(14 dyes)
(~2 8 dyes/target)
(~1020 dyes/target)
(~2040 dyes/target)
(~120200 dyes/target)
Process time
Sensitivity
When to use
106
Best for abundant

targetin samples where
secondary detection may
generate background.

Learn more about
antibody labeling kits at
ablabeling
Best for standard,

everyday immunofluorescence.

Find labeled secondary
antibodies at
antibodies
Best when used with

biotinylated secondary for
moderate amplification, or
with a biotinylated primary
antibody.

Learn more at
streptavidinconjugates
Enzyme-driven signal
amplification is best for
detecting low-abundance
targets.

Learn more at
tsadetection
Cell analysis
Create your own labeled antibodies
We offer a number of Molecular Probes labeling kits for the direct attachment of intensely fluorescent Alexa
Fluor dyes, R-phycoerythrin (R-PE), or even biotin to less than 1 g to up to 1 mg of IgG antibody. Although
the signals from directly labeled antibodies may not be as bright as those observed when using secondary
antibodies, their use eliminates the noise commonly observed when secondary antibodies bind nonspecifically to the sample. Also, directly labeled antibodies allow you to use more than one same-species antibody
in a single staining experiment. The table below will help you choose from our comprehensive selection of
innovative, time-tested, and competitively priced antibody and protein labeling kits that feature streamlined
protocols for labeling and conjugate purification. Go to lifetechologies.com/ablabeling to find more information on all of these antibody labeling options.
Amount of
IgG
Product
Notes
Label
attachment*
Available fluorophores
and labels**
Applications
<120 g
Zenon IgG Labeling

Kits
Antibodies ready to use in 10 min
Noncovalent
Organic dyes, phycobiliproteins
FC, ICC
Covalent
Organic dyes
FC, ICC, IHC
Covalent
Organic dyes
FC, ICC, IHC
Covalent
Organic dyes
FC, ICC, IHC
Covalent
Organic dyes
ICC, IHC, in
vivo imaging,
FC
Covalent
Organic dyes
FC, ICC, IHC
Isotype-specific labeling
Compatible with stabilizing proteins
1020 g
APEX Antibody
Labeling Kits
Antibodies ready to use in 2 hr (~15 min

hands-on time)
10100 g
Small-Scale Protein
Labeling Kits

hands-on time)
Compatible with stabilizing proteins
Optimized for 10150 kDa proteins,

including IgG antibodies (~150 kDa)
Stabilizing proteins must be removed from
sample before labeling
100 g
Monoclonal Antibody
Labeling Kits
Antibodies ready to use in 90 min (~15 min

hands-on time)
Optimized for both monoclonal and polyclonal IgG antibodies
0.53 mg
Antibodies ready to use in 75 min (~10 min

hands-on time)
Includes degree-of-labeling regulator
SAIVI Antibody
Labeling Kits
Uses no organic solvents and produces

azide-free conjugates
1 mg
Large-Scale Protein
Labeling Kits

hands-on time)
Designed to label monoclonal and polyclonal IgG antibodies
FC = flow cytometry; ICC = immunocytochemistry; IHC = immunohistochemistry.

* For additional information on covalent vs. noncovalent labeling, see note and diagram below. ** Organic dyes include fluorophores like Alexa Fluor dyes as well as fluorescein and tetramethylrhodamine. Phycobiliproteins include R-PE, APC, and their tandems (e.g., Alexa Fluor 680R-phycoerythrin (R-PE)).
Covalent vs. noncovalent antibody labeling
Covalent attachment of a label to an antibody generates a stable conjugateone that

lasts several months to years. Labels that are attached noncovalently can dissociate
over timethese conjugates last from several hours up to days.
Noncovalent Zenon antibody labeling (A) vs. covalent
microscale, monoclonal, and protein labeling (B).
107
Cell analysis
Effective labeling of small amounts of IgG
Many IgG antibodies are available only in small quantities and

are often packaged with stabilizing proteins, such as bovine
serum albumin (BSA), or are present in whole serum, ascites,
or complete tissue culture media. These contaminants can
interfere with the labeling reaction. Both the Zenon and
APEX antibody labeling kits provide a convenient means to
label small amounts of IgG antibodies <20 g with methods that
are compatible with these contaminants.
Target-specific
IgG antibody
Zenon labeling reagent

(labeled Fab fragment)
Incubate
Labeled Fab fragments

bind to the IgG
Mix with nonspecific IgG,

which complexes with
unbound Fab fragments
Labeled IgG ready

in only 10 min
Zenon Antibody Labeling Kits

Zenon labeling technology provides a versatile, easy-to-use
system for labeling mouse IgG1, IgG2a, IgG2b, and rabbit IgG
antibodies with our premier Molecular Probes dyes as well as
other fluorophore, biotin, photo-proteins, and enzyme labels.
This new technology offers several advantages over direct
chemical labeling, Including:
Speedentire labeling procedure typically takes 10 minutes

Efficiencylabel nearly 100% of the primary antibody
Economylabel submicrogram amounts of antibody
Simplicityno pre- or post-labeling purification of the anti-
The Zenon labeling scheme. An unlabeled IgG is incubated with the Zenon
labeling reagent, which contains a fluorophore-labeled Fab fragment. The labeled
Fab fragment binds to the Fc portion of the IgG antibody, and the excess Fab fragment is bound by the addition of a nonspecific IgG. The addition of nonspecific
IgG prevents cross-labeling of the Fab fragment in experiments where multiple
primary antibodies of the same type are present. Note that the Fab fragment used
for labeling does not need to be coupled to a fluorophore, but could instead be
coupled to an enzyme or to biotin.
body is required
Flexibilityeasily use multiple primary antibodies in a single

experiment
Learn more at lifetechnologies.com/zenon.
APEX Antibody Labeling Kits
The new APEX Antibody Labeling Kits provide a convenient

means to directly attach a fluorophore to very small amounts
of IgG antibody (1020 g), while allowing you to easily remove
contaminants without losing antibody. APEX Antibody Labeling
Kits utilize a solid-phase labeling technique that captures the
IgG antibody on the resin inside the APEX antibody labeling tip.
This enables covalent labeling of antibodies that are supplied
in solutions containing stabilizing proteins (i.e., BSA) or other
contaminants, which can interfere with the amine-reactive
labeling reagents that are used to attach the fluorophore to
the antibody. With the APEX Antibody Labeling Kit technology,
contaminants are simply eluted through the tip. After applying
the amine-reactive label, the fluorescent IgG conjugate is ready
for use in an imaging (see figure) or flow cytometry application
in as little as 2 hours with very little hands-on time.
Learn more at lifetechnologies.com/apex.
108
Cap
Resin
Tip sheath
APEX Antibody Labeling Kits allow convenient elution of contaminants through

the filter tip.
Cell analysis
Labeled secondary antibodies
Labeled secondary antibodies form stable and specific complexes with unlabeled primary antibodies,
providing the foundation for most immunofluorescence microscopy protocols. Molecular Probes extensive selection of labeled secondary antibodies includes antibodies labeled with our superior Alexa Fluor
dyes, phycobiliproteins, Alexa Fluor dyephycobiliprotein tandem fluorophores, Qdot nanocrystals, biotin,
and enzyme labels (horseradish peroxidase and alkaline phosphatase). We also offer antibodies with
different immunoreactivities, essential to avoid confounding cross-reactivity when performing simultaneous
secondary immunodetection of two or more targets. We also offer F(ab)2 antibodies, which eliminate the Fc
receptor interactions that can be observed when using whole IgG.
Secondary antibody selection guide.

Antibody type
Standard whole IgG
Highly cross-adsorbed whole IgG
F(ab)2 fragment
Adsorbed against
Human IgG and human serum
Bovine IgG, goat IgG, rabbit IgG, rat

IgG, human IgG, and human serum
Human IgG and human serum
When to use
Best for standard, single-color immunofluorescence when cross-reactivity

is not a concern
Best for multicolor immunofluorescence when cross-reactivity may be a

concern
Best for use in immunofluorescence

when Fc interaction observed with
whole antibody interferes with results
Find the labeled antibody youre looking for online
We offer an extensive selection of labeled antibodies and F(ab)2 fragmentsideal for flow cytometry, fluorescence imaging, western blot analysis and more. Use our online Secondary Antibody Selection Tool to
explore a variety of secondary antibodies conjugated to Alexa Fluor dyes, HRP, AP, Qdot nanocrystals,
biotin, and more. You can specify target IgG class (and form), host, species reactivity, and conjugate type to
narrow your results. Find your secondary antibody now at lifetechnologies.com/antibodies.
109
Cell analysis
Choosing the right fluorophore
Browse our broad range of fluorescent labels to find the one that best suits your experiment. We offer organic dyes such as the
Alexa Fluor dyes, phycofluors and their conjugates (often used in flow cytometry), and Qdot nanocrystals, which exhibit exemplary brightness and photostability. Selecting the right fluorescent label requires matching your application and instrument platform to the properties of the dye. The selection guide below in conjunction with our online SpectraViewer can assist you in selecting
the right fluorescence label for your experiment.
Detection reagent
Organic dye
Examples
Alexa Fluor dyes
Texas Red dye
Phycofluors
Qdot nanocrystals
R-phycoerythrin (R-PE) and

conjugates
Qdot 585
Qdot 655
Fluorescein
Allophycocyanin (APC) and conjugates
Photostability
++
+++
Typical analysis platform
FC, FM, HCS
FC
FC, FM
Applications
ICC, IHC
ICC
ICC, IHC, western blot
Brightness
++
+++
+++
Recommended mountant
ProLong Gold antifade reagent,

SlowFade Gold antifade reagent,
PBS
Not applicable
Qmount Qdot mounting medium,

PBS
Online resource
lifetechnologies.com/alexa
streptavidinrpe
lifetechnologies.com/qdot
FC = flow cytometry; FM = fluorescence microscopy; HCS = high-content screening (imaging); ICC = immunocytochemistry; IHC = immunohistochemistry
Improve signal-to-noise ratio with Image-iT

FX Signal Enhancer
Image-iT FX Signal Enhancer (Cat. No. I36933) dramatically

improves the signal-to-noise ratio of immunolabeled cells and
tissues, allowing you to clearly visualize targets that would
normally be indistinguishable. Background staining typically
seen with fluorescent dyes is largely eliminated when ImageiT FX Signal Enhancer is applied to fixed and permeabilized
cells (see figure).
110
Increased label specificity and resolution provided by Image-iT FX Signal

Enhancer. Golgi in fixed and permeabilized HeLa cells labeled with antigolgin-97
(A21270) and visualized with green-fluorescent Alexa Fluor 488 goat antimouse
IgG (A11001). Actin was stained with red-fluorescent Alexa Fluor 594 phalloidin
(A12381); nuclei were stained with blue-fluorescent DAPI (D3571). Cells in the right
panel were treated with Image-iT FX Signal Enhancer (I36933), which eliminates
nonspecific dye binding.
Cell analysis
BackDrop Background Suppressor
BackDrop Background Suppressor (Cat. No. R37603) is a novel reagent from Molecular Probes designed
to effectively suppress background fluorescence in live-cell imaging experiments. BackDrop Background
Suppressor is a direct addition product that comes in ultraconvenient dropper bottles as a six pack with two
vials each of blue, green, and red background suppressors.
Works with fluorescent dyes and proteins

Greatly reduced background fluorescence in live-cell imaging experiments
Higher signal-to-noise, improved sensitivity, and better results
Convenience of direct addition to cells from dropper bottle, no need to remove medium
Flexibility to modulate blue, green, or red background signal
Image-iT Fixation/Permeabilization Kit
Image-iT Fixation/Permeabilization Kit (Cat. No. R37602) is a convenient-to-use fixation/permeabilization

kit that contains all the necessary reagents to prepare your cells for antibody staining and imaging. The highquality components help preserve cell morphology and reduce background staining and come in a practical
box with single-use vials and easy-to-follow protocols.
A complete fixation/permeabilization kit with ready-to-use components and room temperature storage
Optimized for secondary antibody labeling, fixed-cell dye staining, and GFP applications
Convenient storage box, 1X ready-to-use components with 18-month room temperature stability, and
protocol on the inner lid
Compatible with analysis of most cellular antigens

Reduced background staining
Easily accessible online, the Fluorescence SpectraViewer displays the spectral features of one or more fluorophores on a single graph and allows you to overlay representations of your instruments excitation source,
excitation filters, and emission filters as well. This tool simplifies the process of choosing fluorophores that
are compatible with each other in a multicolor experiment and that will also work with your instruments
optics and filters. Find it online at lifetechnologies.com/spectraviewer.
111
Protein analysis
Protein analysis
We have the products to help meet your protein research needs whether you are trying to express proteins in prokaryotic or
mammalian cells, or when you need to verify that the protein you made is the right target. Novex, Ambion, and Invitrogen
brands provide the trusted, reliable, and scalable solutions for your protein research needs.
The chart below highlights the most popular products and new technologies in this chapter. Follow the Web links provided in this
If you have any questions about which of our solutions are right for you, please contact Life Technologies technical support
by phone, email, Facebook at facebook.com/novexprotein, facebook.com/ambion, or facebook.com/invitrogen, or Twitter at
twitter.com/everydayprotein.
Section
Protein analysis
Key
technologies
Protein expression
systems
Transfection
RNAi
Champion pET Expression

System, page 119
Lipofectamine LTX Reagent,

page 124
Silencer Select siRNA, page 126127
Insect expression, page 117
Lipofectamine RNAiMAX
Reagent, page 125
PichiaPink Yeast Expression

System, page 118
GeneArt Algae Engineering Kit,
page 120
GeneArt Gene Synthesis,
page 121
Everyday
essentials
Lipofectamine 2000 Reagent,

page 125
Stealth Select RNAi siRNA, page

126127
mirVana miRNA Mimics and Inhibitors,
page 127128
FreeStyle MAX Transfection

Reagent, page 125
Lipofectamine Reagents,
page 125
siRNA and miRNA, page 126128
Invivofectamine 2.0 Reagent,

page 125
Ambion Pre-miR Precursors and

Anti-miR Inhibitors, page 127128
Silencer siRNA, page 126
Neon Transfection System,

page 124
Instruments
112
Protein analysis
Immunocytochemistry of mouse fibroblasts cells

labeled with AKT [pS473] ABfinity Recombinant
Rabbit Monoclonal Antibody. See page 147 for more
on Novex primary antibodies.
Protein sample prep
Protein analysis
Protein detection
Dynabeads immunoprecipitation, page 129
Bolt gels for western blotting, page

136138
Novex multiplex assays, page 153154
Dynabeads organelle isolation kits,

page 132
Novex multiplex magnetic assays,

page 154155
Sharp and SeeBlue Plus 2 Pre-Stained

Standards, page 141
Novex chemiluminescent ELISAs,

page 151
Protein analysis
PARIS Kit for isolation of total RNA and

protein, page 133
NuPAGE gels, page 136, 138
SYPRO Ruby Protein Gel Stain, page 142

SilverQuest Protein Gel Stain, page 142
iBlot Western Detection Kits, page 145
ABfinity rabbit monoclonal antibodies,
page 147
Protein A/G purification, page 130
Novex Tris-glycine gels, page 136
Novex Antibody Pair kits, page 150
Novex proteases, page 131
Novex primary antibodies for western blotting, page 146
Novex ELISA kits, page 150152
Specialty resins for large and small protein

biomolecules, viruses, and antibodies,
page 131
Novex cell lysis reagents, page 133
POROS Analytical Chromotography,

page 131
ZOOM IEF Fractionator, page 132
SeeBlue Pre-Stained Standard, page 141

SimplyBlue SafeStain, page 142
Qubit 2.0 Fluorometer, page 134
Luminex MAGPIX System, page 155
XCell SureLock protein electrophoresis

cells, page 139
Luminex 200 System, page 155
iBlot 7-minute blotting system, page 143
FLEXMAP 3D High-Throughput System,

page 155
BenchPro 4100 Western Processing

System, page 144
113
Protein analysis
Protein expression systems

Recombinant protein expression technology enables analysis of gene regulation and protein structure and function. Utilization
of recombinant protein expression varies widelyfrom investigation of function in vivo to large-scale production for structural
studies and biotherapeutics drug discovery. Using the right expression system for your specific application is key to your
success. Protein solubility, functionality, speed, and yield are often the most important factors to consider when choosing an
expression system. With the wide variety of expression systems available from Life Technologies, youre sure to find one that
meets your needs. The following table summarizes some of the characteristics of the most popular expression hosts. Several
expression systems from Life Technologies are detailed on the following pages. For a complete list of expression systems, visit
lifetechnologies.com/proteinexpression.
Expression hosts and their applications

Host organism
Most common applications
Advantages
Challenges
Cell-free prokaryotic
Rapid expression screening
Rapid expression directly

from plasmid
Large-scale expression
(3 mg)
Toxic proteins
Incorporation of unnatural
labels or amino acids
Functional assays
Protein expression
Protein interactions
Open systemeasily add

components to enhance
solubility/functionality
Overall cost
and time
Lower
Simple format
Scalable
Compatible with Gateway
cloning
Prokaryotic
Yeast
Structural analysis
Scalable
Protein solubility
Antibody generation
Low cost
Functional assays
Simple culture conditions
Minimal posttranslational
modifications

cloning
May be difficult to express

functional mammalian
proteins
Structural analysis
Eukaryotic protein
processing
Fermentation required for

very high yields
Scalable up to fermentation
(grams/liter)
Growth conditions may

require optimization
Antibody generation
Functional assays
Insect
Functional assays
Structural analysis
Antibody generation
Simple media requirements

Posttranslational modifications similar to mammalian
systems
More demanding culture

conditions
Greater yield than mammalian systems

cloning
Mammalian
Functional assays
Antibody generation
Highest level of correct posttranslational modifications

Highest probability of
obtaining fully functional
human proteins
cloning
114
Multimilligram/liter yields
only possible in suspension
cultures
More demanding culture
conditions
Higher
Probability of
obtaining functional
protein
Protein analysis
Protein expression custom services
Let the experts help you express your protein at the quality and quantity you need
Expression optimization
Protein production and scale-up
Protein purification
Life Technologies offers a variety of protein expression, optimization and purification services tailored to meet
your individual needs. Our experienced, highly trained scientists can work with you to select the expression
systems that best suit your applications and then perform all the steps necessary for obtaining high-quality
preparations of your protein of interest. For additional information or price quotations, contact us via phone
at 800 955 6288 x45682 (in North America), via email at custom.services@lifetech.com, or visit us online at
lifetechnologies.com/customservices.
Selecting a mammalian expression system
There are many options in selecting a mammalian expression system matched to your specific needs. Ask
yourself the three following questions, then use the table below to match the best system to meet your needs.
1. How much protein do you need?
Nanograms to micrograms
Milligrams to grams
Protein expression
Protein yields are highly variable and depend largely on the specific protein to be expressed. In general, small
amounts of protein can be readily generated using transient transfection of plasmid DNA into a wide variety
of cells. It is very important to achieve the highest transfection efficiency possible using effective transfection
reagents such as Lipofectamine LTX or Lipofectamine 2000 reagents.
To produce larger amounts of protein (milligrams to grams) requires more cells. Thus, stable cell lines
are often employed to produce a large, selected population of cells that is stably expressing your protein of
interest. Alternatively, large-scale transient transfection of suspension-adapted cells such as HEK 293 or
CHO cells can generate large amounts of protein in a much shorter time period than can be achieved with
stable cells. See the following section to understand the choices for producing large amounts of protein.
2. How quickly do you need to obtain your protein?
Scalable transient expression delivers high quantities of protein in a short period of time
Stable cell lines produce the highest amounts of protein but require a longer time to set up
Expression by transient transfection results in high levels of expression for a few days to a week. Thus its
ideal for rapid protein production and quick data generation. Transient expression systems such as the FreeStyle 293, FreeStyle MAX CHO, and Expi293 Expression System use suspension-adapted 293 or CHO
cells at culture scales from 1 mL to 100 L to generate large amounts of protein. The new Expi293 Expression System has been demonstrated to produce up to 1 gram of protein per liter of cell culture.
To produce a cell line that can be used over a long experimental time course or over many experiments,
it is necessary to generate a stable cell line in which your expression construct is integrated into the host
genome. A selection agent, added to the medium, is used to select the cells that have integrated the
construct. If you want to make cell lines with targeted integration into an expression hotspot, we recommend the Jump-In System. For randomly integrated stable cell lines, any pcDNA vector delivered by
standard cationic transfection reagent, or the Neon Transfection System, or the ViraPower Lentiviral
Expression System may be used.
115
Protein analysis
3. Is it important for you to control when expression begins?
Expression systems with constitutive promoters do not allow you to control expression
Expression systems with inducible promoters require you to add an inducer to begin expression
If you are working with a nontoxic gene and the timing of expression is not important, choose a constitutively
expressing promoter.
An inducible promoter allows you to control the timing of gene expression. In the absence of an inducer, your
gene is not expressed. Add inducer to turn on expression. This option is ideal for expressing toxic proteins.
Choose from the T-REx Inducible Expression System or the Flp-In T-REx System. For more information on inducible promoters, please visit lifetechnologies.com/mammalianexpression.
Application
System
Key features
Constitutive expression (CMV

promoter)
pcDNA vectors
Constitutive CMV expression with your choice of

several epitope tags and selection markers
Transient expression
pcDNA vectors and cationic transfection reagents

Adenoviral Expression Systems
High-level gene expression in any dividing or nondividing mammalian cell type
Scalable, transient expression
FreeStyle 293 Expression System

FreeStyle MAX CHO Expression System
Expi293 Expression system
Scalable suspension culture transfections from

1 mL to 100 L
Obtain protein in several days to 1 week
Protein expression
Production of up to 1 gram of protein per liter of

culture
Stable cell lines, targeted
integration
Inducible expression
Jump-In Fast Gateway System
Rapid generation of stable cell lines with integration

into expression hotspots
Jump-In TI-Gateway System
Generation of stable cell lines with integration into

preselected genomic sites
Flp-In T-REx System
Regulated expression from CMV promoter
T-REx System, Flp-In T-REx System
Rapid generation of regulated stable expression cell

lines
ViraPower Lentiviral T-Rex System
Regulated expression in any dividing or nondividing

mammalian cell type
Find out more about mammalian expression systems at lifetechnologies.com/mammalianexpression.
116
Protein analysis
Insect expression systems

High yields, more functional proteins
Expression in insect cells offers significant advantages, including high expression levels, ease of scale-up,
production of proteins with proper posttranslational modifications, and simplified cell growth. Insect cells
do not require CO2 for growth and can be readily adapted to high-density suspension culture for large-scale
expression. Many of the posttranslational modification pathways present in mammalian systems are also
utilized in insect cells, allowing the production of recombinant protein that is antigenically, immunogenically,
and functionally similar to the native mammalian protein. Baculovirus expression systems are powerful
and versatile systems for high-level recombinant protein expression in insect cells. Expression levels up to
500 mg/L have been reported using the baculovirus expression system.
The BaculoDirect Baculovirus Expression System is the fastest and easiest method available for
Protein expression
generating recombinant baculovirus. The BaculoDirect Linear DNA includes attR sites for rapid and
efficient recombinational cloning with a Gateway entry clone. The resulting recombinant DNA is taken
directly from the LR reaction mix and used to transfect insect cells, which helps to save a significant
amount of time. Purified virus can typically be isolated within one week. The reduction of hands-on time
makes BaculoDirect ideal for high-throughput expression.
The Bac-to-Bac Baculovirus Expression System uses a unique bacmid shuttle vector that recombines
by site-specific transposition and generates an expression bacmid in bacterial cells. The expression
bacmid is then transfected into insect cells to generate recombinant baculovirus.
The Bac-N-Blue Baculovirus Expression System has been used for over a decade to produce high
levels of recombinant proteins.
The Drosophila Expression System (DES) uses the well-characterized Drosophila Schneider S2 cells and
simple expression vectors to allow stable or transient expression of recombinant proteins.
Selection guide for insect expression systems.

Fusion partner
Position
Purif.
Epitope
Promoter
Expression/
inducer
N-term
C-term
6xHis
6xHis
V5
V5
Polyhedrin
Infection
Fast and easy method

for generation of
recombinant baculovirus;
ideal for high throughput
Honeybee
melittin
GST
N-term
6xHis
pFastBacHT
pDEST10
Polyhedrin
or p10
Infection
production
Rapid baculovirus
production; easy blue/
white selection of
recombinant colonies
Sf9, Sf21,
or High
Five
Honeybee
melittin
C-term
6xHis
Xpress V5
Polyhedrin
Infection
Classic and trusted

expression system for
high-level recombinant
protein production
S2 cells
BIP
C-term
6xHis
V5
MT or Ac5
CuSO4 or
constitutive
Easy-to-use stable
system, constitutive or
inducible expression;
uses simple plasmids for
expression; extremely
high integration of
transfected plasmids
eliminates need to select
and screen for expression
from clonal cell lines
System
Host
BaculoDirect
Sf9, Sf21,
or High
Five
Bac-to-Bac or
Bac-to-Bac HBM
Sf9, Sf21,
or High
Five
Bac-N-Blue
DES
Secretion
signal
Advantage
Find out more about insect expression systems at lifetechnologies.com/insectexpression.
117
Protein analysis
PichiaPink Yeast
Expression System
Large-scale production of secreted

recombinant proteins
Screen Pichia transformants rapidly by adenine auxotrophy and color selection
Reduce protein degradation with protease-deficient Pichia pastoris strains
Optimize protein secretion with one of eight secretion signal sequences
The PichiaPink System is a eukaryotic protein expression system based on the
eukaryote Pichia pastoris, which can be used for high-level (grams per liter) and largescale (>1,000 L) production of secreted recombinant proteins. This system contains both
low- and high-copy plasmid backbones, eight secretion signal sequences, and four
yeast strains to help you optimize for the highest possible yield of recombinant protein.
Protein expression
P. pastoris carries out many of the same posttranslational modifications that occur in
mammalian cells, which means that the likelihood of producing a functional recombinant protein from this expression system is very high. Additional advantages of
P. pastoris include rapid growth, a well-defined genetic background, simple media
formulations, and simple handling techniquesall of which make it the ideal host for
producing large quantities of protein in a short period of time.
Use color selection to find your transformed Pichia

Pink colonies. Transformed Pichia appear as white
colonies, while untransformed Pichia appear as redpink colonies.
The PichiaPink System provides you with two different kits for secreted expression
of your recombinant protein of interest:
The PichiaPink Secretion Optimization Kit enables you to screen multiple signal
sequences with your gene of interest in both low- and high-copy vectors (pPink-LC
and pPink-HC, respectively) for optimal expression and secretion of your recombinant protein. pPink-LC and pPink-HC vectors are also available separately as the
PichiaPink Vector Kit.
The PichiaPink Secreted Protein Kit allows you to clone your gene interest in
frame with the Saccharomyces cerevisiae -mating factor presequence using the
pPink-HC plasmid for secreted expression of your recombinant protein. pPink-HC
is also available separately as the PichiaPink Secreted Protein Vector Kit.
PichiaPink Secretion Optimization
Kit contents:
PichiaPink Secreted Protein

Kit contents:
The PichiaPink Vector kit containing
The PichiaPink Secreted Protein
a high- and low-copy plasmid

The PichiaPink Secretion Signal set
containing 8secretion signals
The PichiaPink Expression Strain
set containing 4glycerol strains
The PichiaPink Media Kit containing
4 basic media
Vector kit containing the highcopy plasmid with the S. cerevisiae

-mating factor presequence to yield
secreted protein expression
The PichiaPink Expression Strain
set containing 4glycerol strains
The PichiaPink Media Kit containing
4 basic media
Find out more about yeast expression systems

at lifetechnologies.com/yeastexpression.
118
Protein analysis
Prokaryotic expression systems
Easy-to-use recombinant protein expression systems

Due to their ease of use, relatively high yields, and simple scalability, T7-regulated E. coli expression systems
are often chosen for generating recombinant protein. Selecting the right system for your specific application
is the key to your success. With the wide variety of T7 expression systems available from Life Technologies,
youre sure to find one that meets your needs. The table below summarizes some of the most popular T7
expression systems. For a complete list, go to lifetechnologies.com/proteinexpression.
Champion pET Expression Systems offer the high-level expression

Protein of interest constitutes >50% of total cellular protein
5-minute TOPO Cloning with >95% efficiency
No ligase, post-PCR procedures, or restriction enzymes required
Protein expression
The Champion pET Expression System uses optimized vectors and a BL21 Star E. coli expression strain to
produce protein yields up to 10-fold greater than any other expression system. The system takes advantage of
the high activity and specificity of the bacteriophage T7 RNA polymerase for high-level transcription of the gene
of interest. The lac operator located in the promoter region provides tighter regulation than traditional T7-based
vectors, improving plasmid stability and cell viability. BL21 Star E. coli are genetically engineered to reduce
transcript degradation, resulting in significantly improved mRNA stability and increased protein production.
Cloning into a Champion pET Expression vector is easy, fast, and efficient. Available with Directional
TOPO or Gateway Cloning technologies, you can typically generate your clone in one day. In addition,
the Champion pET vectors offer simplified protein purification, protein detection, enhanced solubility, and
native protein expression.
Selection guide for our most popular T7 expression systems.

System
Promoter
Advantage
Champion pET302/NT-His and pET303/CT-His Vector Kit
T7 lac
Higher expression levels than those obtained from other pET vector
suppliers
Champion pET300/NT-DEST and pET301/CT-DEST

Gateway Vector Kit
T7 lac
Convenience of Gateway cloning and choice of N-term or C-term

6xHis tag
Champion pET Expression System
T7 lac
Highest level of protein production
Champion pET SUMO Expression System
T7 lac
Highest level expression of proteins with enhanced solubility
Champion pET Expression System
T7 lac
Rapid, visual screening of protein expression without western

blotting with Lumio technology
Champion pET BioEase Expression System
T7 lac
Easy, efficient expression of biotinylated proteins
T7 Expression System
T7
High-level expression of nontoxic proteins
pBAD Expression System
araBAD
Tightly regulated expression for efficient expression of difficult-toexpress proteins
trc Expression System
trc
Enhanced expression of eukaryotic protein
Find out more about prokaryotic expression systems at lifetechnologies.com/bacterialexpression.
119
Protein analysis
GeneArt Algae Engineering Kits

Genetic modification and expression systems for
photosynthetic microalgae
GeneArt Algae Engineering Kits for Chlamydomonas reinhardtii and Synechococcus elongatus are the first
commercially available genetic modification and expression systems for photosynthetic microalgae. These
kits are designed for rapid scale-up and production and consistent, defined quality.
Algal cells arrive ready to resuscitate, grow, and transform or store at 80C until ready to use
Every cell lot is manufactured using a standardized manufacturing protocol, so every experiment starts
with consistent materials
Optimized media, vectors, and protocols help ensure robust selection and expression
Get transformed cells more quickly
GeneArt Algae Engineering Kits save time required for strain optimization and transitioning to downstream
applications such as bioproduction.
Protein expression
Cells are typically ready for transformation in less than 5 days

GeneArt vectors facilitate rapid directional cloning of synthetic genes or PCR products
Gibco media are provided at 1X concentration, eliminating time-consuming media preparation
Take control of the quality of your reagents
Dont wonder about the condition and characteristics of your cultures and expression clones. With GeneArt
Algae Engineering Kits, you have access to a quality-controlled supply of algae cells and cloning tools.
Kits are shipped and stored at 80C

Algal cell stocks are viable, pure, and standardized
GeneArt vectors are fully characterized and optimized for engineering and expression in each strain
Find out more about algae expression systems at lifetechnologies.com/algaeexpression.
Thaw and resuscitate algae (24 days)
50
45
40
35
30
25
Grow algae in light

for 24 days to
OD >0.6
750
Transform algae and test for expression (56 days)
20
15
10
7.5
5
Construct and verify the expression clone (3 days)
Amplify the gene of

interest, insert into
vector, and confirm
GeneArt Algae Engineering Kits offer quality-controlled components and significant process efficiencies for both research and bioproduc
tion environments. Following the optimized workflow, you can typically see transformed cells in less than 10 days.
120
Protein analysis
GeneArt gene synthesis service
Let us synthesize your DNA constructs for improved or

engineered protein expression
Gene synthesis has become the most cost-effective, time- and resource-saving method for obtaining nearly
any desired DNA construct, producing expression clones that outperform those produced using conventional
molecular biology techniques in expression performance, stability, and quality.
GeneArt gene synthesis tools go beyond traditional synthesis and enable you to:
Improve protein expression with proprietary GeneOptimizer technology

Gain access to hard-to-clone constructs, long, complex DNAs, and customized vectors
Create unlimited numbers of mutants for screening experiments
Overcome RNAi inactivation
Engineer proteins to improve enzymes and humanize and/or increase binding affinities of antibodies
Find out more about gene synthesis at lifetechnologies.com/genesynthesis.
www
G
A
A
G
Protein expression
5 5 5 5 5 5 5 5 5 5 5 5 5 5
A C G C A
Day 1
Ordering until
3:00 pm (CET)
Oligosynthesis
over night
Day 2
Gene Assembler
Day 3
Cloning
Day 4
Sequencing &
quality control
Day 5
Ready for shipment
GeneArt SuperSPEED production schedule.
GeneArt GeneOptimizer sequence optimization
Our proprietary GeneOptimizer algorithm offers proven increases in protein yield through sequence optimization. By evaluating many important expression parameters in parallel, GeneOptimizer technology
generates up to 500,000 variants of your target sequence in an evolutionary approach and selects the best
match for your specific requirements. Sequence optimization using the GeneOptimizer software process is
included as an optional step with all GeneArt gene synthesis services.
Select GeneOptimizer technology when you need:
Removal of introns
Knockout of cryptic splice sites and RNA destabilizing sequence elements
Increased RNA stability
Adaptation of codon usage
Extensive mutagenesis
Flexible combination of functional domains
Introduction of restriction sites
Epitope shuffling
Consideration of immunomodulatory CpG motifs
Find out more about gene optimization at lifetechnologies.com/geneoptimization.
121
Remove sequence
repeats
PABP
Adjust codon usage
ng Pr55/g total protein
Optimize GC content
PABP
PABP
AAAAAA
AAAAAA
Eliminate killer motifs
Remove splice sites
3,5
M
oc
k
ty
pe
ild
W
Capture assay
Op
tim
iz
e
ty
pe
Op
tim
iz
ed
ild
W
M
oc
k
Protein analysis
Northern
3
2,5
2
gag
1,5
1
0,5
0
Western
Avoid RNA secondary

structure
Optimized gene
Beta-actin
Sequence optimization using the GeneArt GeneOptimizer process significantly

increases expression. Reference: J Virol 74:10822 (2000).
Proprietary GeneOptimizer algorithm determines optimal gene sequence for

your experiments.
Protein expression
Selection guide for protein expression clones and expression optimization services.
Order now, express in a week

100% sequence-verified clone
available in less than a week
Configure your projectprotein

expression with no gene design
constraints
Optimize your expression

improved protein expression
levels through gene optimization
Product name
Human Ultimate ORF Clones
GeneArt gene synthesis
GeneArt GeneOptimizer
sequence optimization
Cat. No.
https://orf.invitrogen.com/
cgi-bin/ORF_Browser
817003DE (0.53 kb, online

ordering)
817000DE (<0.5 kb, online
ordering)
817003DE (0.53 kb, online

ordering)
817000DE (<0.5 kb, online
ordering)
Key features
Fastest way to obtain a

sequence verified clone
Full service that delivers validated

genes with no limitation in
sequence design
Full service that delivers validated

genes, with improved expression
yields and no limitation in
sequence design
Time to delivery
1 week
615 days to receive ready-to-use

clones
615 days to receive ready-to-use

clones
Gene Size
One gene
Up to 20 kb
Up to 20 kb
Number of fragments
NA
NA
What is delivered
Glycerol stock of uORF entry

clone
Synthetic wild type gene
Optimized gene
5 g lyophilized DNA cloned in

GeneArt standard vector
5 g lyophilized DNA cloned in

GeneArt standard vector
Quality assurance
documentation, including
sequence verification
Quality assurance
documentation including
sequence verification
ORF card data available online
Choice of expression vectors
122
Any Gateway destination vector Any vector
Any vector
Improvement of expression yields NA
NA
Up to 100-fold
Free web-based tool for search

and design
Yes
Yes
Yes
Protein analysis
Overview of transfection
Life Technologies reagent-mediated and electroporation-based transfection products are the most cited and trusted in the industry.
Regardless of your cell type, throughput requirements, or budget, we have the right solution for you.
Fits your experimentwhether youre transfecting plasmid DNA, siRNA, mRNA, miRNA, RNAi plasmids, or protein expression
constructs, we have the right product for you
Easy to useall of our products come with clear and simple protocols for maximum convenience and ease of use
Works with your cellsour transfection systems and reagents were designed for efficient transfection of a broad range of cell
types, including adherent, suspension, and difficult-to-transfect cells
Used by scientists worldwideLife Technologies is the most cited and trusted supplier of transfection reagents, with over
48,000 citations for the Lipofectamine brand alone
The table below outlines our most popular transfection products. If you go to lifetechnologies.com/transfection, you can find information about our entire line of transfection products and youll also have access to:
Transfection and RNAi
Selection guides and performance data for choosing the correct transfection product
Product tables sorted by sample type and transfection method
Cell typespecific transfection protocols for more than 120 cell lines
FAQs and information on selection antibiotics to help you prepare for your experiment
Selection guide for highlighted transfection reagents and systems.
Find this interactive table online at lifetechnologies.com/lipofectamine.
Value reagent for
common cell types
Most cited and versatile

reagent for a wide range of
cell types
Most efficient
reagent for plasmid
delivery; potent and
low cytotoxicity
Most efficient
reagent for siRNA/
miRNA delivery;
efficient gene
knockdown
High-efficiency
electroporation
for all cell lines
Lipofectamine
Lipofectamine 2000
Lipofectamine LTX
Lipofectamine
RNAiMAX
Neon
Transfection
System
Top-selling format
Cat. No. 18324012
Cat. No. 11668019
Cat. No. 15338100
Cat. No. 13778075
Cat. No.
MPK5000S
Sample type
Plasmid DNA
Plasmid DNA
Plasmid DNA
RNAi plasmids
RNA (e.g., siRNA,

miRNA)
Plasmid DNA
RNA (e.g., siRNA, miRNA)

RNAi plasmids
RNA (e.g.,
siRNA, miRNA)
Protein
RNAi plasmids
Transfection
efficiency for
common cell types
Best for primary and
stem cells
Standard
High
Superior
Superior
Maximal
Watch the video (lifetechnologies.com/tfinsights) to learn about the transfection research program and key
factors for successful transfections, which resulted in enhanced protocols for the Lipofectamine line of
transfection reagents.
123
Protein analysis
Neon Transfection System
Superior transfection efficiency for labs of all sizes, budgets, and needs
Maximum efficiencyup to 90% in many cell types, including difficult-to-transfect

cells, primary, and stem cells
Flexibleeasily transfect from 1 x 104 cells to 5 x 106 cells per reaction with easyto-use protocols
Simpleeasy to use, with single reagent kit for all cell types
Versatileopen system allows electroporation parameters to be optimized freely
Learn more about the advantages of the Neon Transfection System
at lifetechnologies.com/neon.
B
High transfection efficiency of Jurkat cells with the Neon Tranfection System. Jurkat cells were transfected
with an EGFP-encoding vector using the Neon Transfection System and viewed using (A) bright-field microscopy
and (B) fluorescence microscopy 24 hr following transfection.
Lipofectamine LTX and PLUS Reagent
Efficient reagent for plasmid delivery and protein expression
Superior performancehigh transfection efficiency with maximal (>90%) cell

viability
Easysimple protocol provided for many cell lines

Versatilesuperior plasmid delivery into a broad range of cell types including
primary and hard-to-transfect cells
Learn more about the advantages of Lipofectamine LTX Reagent at
lifetechnologies.com/ltx.
0 ng
500 ng
1,000 ng
Lipofectamine LTX Reagent with PLUS Reagent, efficiently transfects primary neural progenitor cells.
Lipofectamine LTX Reagent (1.5 L) was used to transfect murine embryonic primary neural progenitor cells with
the indicated quantities of a GFP-expressing plasmid in the presence of PLUS Reagent (2.5 L). GFP expression
was analyzed 24 hr post-transfection. Data courtesy of the Beverly Davidson laboratory, University of Iowa.
124
Protein analysis
Lipofectamine RNAiMAX Reagent
Efficient reagent for siRNA or miRNA delivery in vitro, potent gene knockdown
Superior performanceexcellent transfection efficiency and gene knockdown

Easysimple protocol, minimal cytotoxicity across a 10-fold concentration range of transfection reagent
Versatiledeliver siRNA or miRNA into a broad range of cell types including primary and hard-totransfect cells
Learn more about the advantages of Lipofectamine RNAiMAX Reagent at lifetechnologies.com/rnaimax.
Lipofectamine 2000 Reagent
Most cited and versatile reagent for a broad range of cell types, co-transfection
Provenmost cited transfection reagent with exceptional transfection efficiency

Easysimple protocol, efficiency in the presence of serumeliminates need to change media following
transfection in most cases
Versatileeffective for both plasmid and siRNA or miRNA delivery and co-transfection
Learn more about the advantages of Lipofectamine 2000 Reagent at lifetechnologies.com/lf2000.
Lipofectamine Reagent
Value reagent for common cell types
Reliableappropriate for establishing stable cell lines

Flexibleproven to work in high-throughput applications
Effectiveworks well with PLUS reagent for higher protein expression
FreeStyle MAX Transfection Reagent
Animal origin-free reagent for transient transfection of CHO and HEK 293 cells
High yielddesigned for recombinant protein bioproduction applications

Efficientdesigned for the highest expression levels and transfection rates with the lowest cytotoxicity
Rapidup to milligram quantities (including all posttranslational modifications) produced in one week
Find out more at lifetechnologies.com/freestyle.
Invivofectamine 2.0 Reagent
Easy-to-use, effective reagent for siRNA or miRNA delivery in vivo
Effectivespecific, effective knockdown (>80%) in mouse liver

Simpleeasy-to-use procedure; just mix, equilibrate, and inject
Nontoxic and non-immunostimulatory
Find out more at lifetechnologies.com/invivofectamine.
Meet the Inventor: Invivofectamine 2.0 Reagent

Watch this video (lifetechnologies.com/inventorinvivo) to learn more about Invivofectamine 2.0 Reagent,
the latest innovation for in vivo siRNA delivery from Life Technologies.
125
Protein analysis
Overview of RNAi
Two types of small RNA molecules function in RNAi. The first type comprises synthetic, short interfering RNA (siRNA) molecules
that target mRNA cleavage effectively knocking down expression of a gene of interest. The second are microRNA (miRNA) molecules
that regulate gene expression by binding to the 3 untranslated regions (UTRs) of target mRNAs to inhibit their function. There are
several ways to induce RNAi: synthetic molecules, RNAi vectors, and in vitro dicing. Your choice of tools depends on your model
system, the length of time you require knockdown, and many other experimental parameters.
The sections below highlight some of our most popular RNAi products, but you can view even more at lifetechnologies.com/rnai.
There youll find:
Guides for choosing the correct RNAi delivery method for your experiment
The 160-page RNAi Sourcebook (downloadable PDF)
Easy-to-use online ordering tools for custom RNAi synthetics and vectors and custom miRNA inhibitors and precursors
Made-to-order and preplated siRNA libraries
siRNA positive and negative controls
Protocols and guidelines to help you plan and execute your experiment
For vector-based RNAi products, visit lifetechnologies.com/vectorrnai.
siRNA
Silencer Select siRNA

Highest knockdown, lowest off-target effects
100-fold more potent knockdown
Lowest off-target effects via locked nucleic acid (LNA)
chemistry
100% guaranteed to silence (>70%) (terms and conditions
apply; go to lifetechnologies.com/silencerselect)
Stealth Select RNAi siRNA
Good knockdown, low off-target effects
High knockdown
Low off-target effects
100
94%
90
70
60
50
86%
87%
68%
% effective
80
% mRNA remaining
Traditional RNAi methods for gene knockdown in mammalian

cells involved the use of synthetic RNA duplexes consisting of
two unmodified 21-mer oligonucleotides annealed together to
form short/small interfering RNAs (siRNAs). Silencer Select
siRNA and Stealth RNAi siRNA improve upon these traditional
duplexes by using proprietary chemical modifications to ensure
better RNAi results.
70%
Knockdown
Silencer siRNA
Designs
80%
Knockdown
Silencer Select
siRNA Designs
40
30
20
10
0
1 6 11 16 21 26 31 36 41 46 51 56 61 66 71 76 81 86 91 96 101 106 111 116 121 126 131 136 141 146 151
Silencer Select siRNA design algorithm significantly improves effective siRNA

prediction accuracy. The Silencer Select siRNA design algorithm was used to
design 155 siRNAs to 40 different targets. These siRNAs were tested side by side
with siRNAs designed using the previous algorithm at 5 nM in HeLa cells. mRNA
knockdown was measured 48 hr post-transfection via qRT-PCR using TaqMan
Gene Expression Assays. Results are expressed as percent of mRNA remaining
compared to Silencer Negative Control #1 siRNAtreated cells. The inset shows
the percentage of siRNAs that elicited 70% and 80% mRNA knockdown.
Silencer siRNA
Reliable, affordable siRNA
Cost-effective
Good target specificity
126
Protein analysis
Which siRNA is right for you? Find this interactive table online at lifetechnologies.com/siRNA.
Cost-effective siRNA
Good knockdown, low off-target
Highest knockdown, lowest off-target
Silencer siRNA
Stealth Select RNAi siRNA
Silencer Select siRNA
Potency
100 nM
recommended conc.
20 nM
recommended conc.
5 nM
recommended conc.
Efficacy (>70% knockdown)
2 of 3 siRNA guaranteed
Target specificity
Moderate
High
Highest
Innate immune response
Minimum
Minimized through chemical

modifications
Minimized through chemical

modifications
Molecular format
Unmodified 21 bp duplex with

overhangs
Modified 25 bp duplex with no

overhangs
LNA-modified 21 bp duplex with

overhangs
Coverage
Coding RNA
Coding RNA
Coding and noncoding RNA
Target species
Human, mouse, rat (use custom

tool for other species)
Human, mouse, rat (use custom

tool for other species)
Human, mouse, rat (use custom tool for

other species)
Recommended product
Silencer Pre-designed siRNA,

5 nmol (Cat. No. AM16708)
Stealth RNAi siRNA Tube

(Cat. No. 1299003)
Silencer Select Pre-designed siRNA,

5 nmol (Cat. No. 4392420)
miRNA
MicroRNAs (miRNAs) are short, highly conserved small noncoding RNA molecules naturally occurring in
the genomes of plants and animals. miRNAs are 1727 nucleotides long and regulate posttranscriptional
mRNA expression, typically by binding to the 3 untranslated region (3-UTR) of the complementary mRNA
sequence, resulting in translational repression and gene silencing. Studies have shown that thousands of
human protein-coding genes are regulated by miRNAs, indicating that miRNAs are master regulators of
many important biological processes.
Since the discovery of the first lin-4 miRNA in C. elegans, miRNAs have been identified in diverse organisms
through experimental determination or computational prediction. These are managed through the miRBase
database, which is a searchable public database of all published miRNA sequences and annotations.
mirVana miRNA Mimics and Inhibitors
Next-generation miRNA
Low off-target effects and high potency
100% coverage of miRBase v 17
In vitro and in vivo use
Ambion Pre-miR Precursors and Anti-miR Inhibitors
Reliable, affordable miRNA
High specificity and efficient inhibition
100% coverage of miRBase v 15
Meet the Inventor: Silencer Select siRNA

Susan Magdaleno describes Silencer Select siRNA technology
(watch the video at lifetechnologies.com/inventorsirna).
127
Protein analysis
1.20
450
FW
RV
Fold change
1.00
0.80
0.60
0.40
0.20
350
300
250
200
150
100
50
0
0.00
miRNA
mimic
Neg
reporter 1
miRNA
mimic
Neg
miRNA
mimic
reporter 2
Neg
reporter 3
miRNA
mimic
Neg
miRNA
mimic
reporter 4
Neg
reporter 5
miRNA
mimic
Neg
miR
122
reporter 6
mirVana miRNA mimic mature strand is highly potent while star strand is inac
tivated. The key advantage of mirVana miRNA mimics is inactivation of the star
strand. miRNA mimics, like natural microRNAs have 2 strandsthe mature strand
(guide strand) that is functional and used by Ago protein to target mRNAs; and
the star or passenger strand, which is nonfunctional and is normally cleaved and
expelled from the complex. Most scientists want to analyze one strand of miRNA at
a time, and want the other strand to be totally inactive: mirVana miRNA mimics
achieve this. For this assay we measured activity from both strands of miRNA
mimics. One reporter has a target in forward orientation to measure activity of the
mature miRNA strand; another reporter has the target cloned in reverse/complement orientation to test activity of the star strand of the miRNA mimic. For all 6
sequences, activity of the mature strand is high (5- to 10-fold lower than negative
control), and activity of the star strand is low or nothing (similar to neg control).
miR122 = miR122 miRNA inhibitor

NT = Not transfected
Neg1 = Negative Control #1
400
mRNA upregulation %
1.40
NT
AldoA
Neg1
miR
122
NT
Neg1
Hfe2
miR
122
NT
Neg1
Slc35a4
miR
122
NT
Neg1
Lass6
mirVana miRNA Inhibitors effectively suppress miRNA in vivo. miR122 or Negative

Control #1 mirVana miRNA inhibitors were complexed with Invivofectamine 2.0
Reagent and delivered to Balb-C mouse liver via tail vein injection on three consecutive days at a dose of 7 mg/kg body weight. Expression levels of four mRNA targets
(AldoA, Hfe2, Slc35a4, and Lass6), natural targets of miR122, were measured with
TaqMan Assays in the livers of mice injected with miR122 inhibitor, Negative Contol
#1 (Neg 1) as well as untreated animals. Significant up-regulation of mRNA was
detected in livers of mice treated with miR122, compared to untreated and negative
control-treated mice; >3.5-fold for AldoA, >2-fold for Hfe2, >3-fold for Slc35a4, and
>4-fold for Lass6. This indicates that mirVana miRNA inhibitors are efficiently
delivered to the liver with Invivofectamine 2.0 Reagent and inactivate miR122,
leading to up-regulation of genes naturally suppressed by miR122.
Which miRNA mimic or inhibitor is right for you? Find this interactive table online at lifetechnologies.com/miRNA.
Novel design to minimize
off-target effects
Next-generation chemistries Chemically modified for

for lowest off-target effects
good efficacy
Next-generation chemistries
for highest efficacy
Ambion Pre-miR
Precursors
mirVana miRNA
Mimics
mirVana miRNA
Inhibitors
Ambion
Anti-miR Inhibitors
Function
Mimic endogenous miRNAs
Mimic endogenous miRNAs
Inhibit endogenous miRNAs
Inhibit endogenous miRNAs
Content
database
100% coverage of miRBase

v 15*

v 17**

v 15*

v 17**
Model system
In vitro
In vitro and in vivo
In vitro
In vitro and in vivo
Species covered
Human
Human, mouse, rat (use

Human
custom tool for other species)
Human, mouse, rat (use

custom tool for other species)
Custom design
tool
Design your custom miRNA

mimics

mimics

inhibitors

inhibitors
miRNA libraries
Predesigned or custom
libraries
libraries
libraries
libraries
Recommended
product
Pre-miR miRNA Precursor,

5 nM (Cat. No. AM17100)
mirVana miRNA Mimic,

5 nM (Cat. No. 4464066)
Anti-miR Inhibitor,
5 nM (Cat. No. AM17000)
mirVana miRNA Inhibitor,

5 nM (Cat. No. 4464084)
* 1,090 distinct human sequences.

** 1,719 distinct human sequences.
Meet the Inventor: mirVana mimics and inhibitors

Alexander (Sasha) Vlassov describes mirVana miRNA mimics and inhibitors and their applications
for functional analysis (watch the video at lifetechnologies.com/inventormirna).
128
Protein analysis
Protein sample preparation

Life Technologies offers reliable solutions to isolate peptides, proteins and protein complexes directly from your crude sample for
further analysis to advance your proteomic workflows.
Select the protein sample preparation method for your sample using the table below. Further information on each of the recommended methods can be found in the pages that follow.
Protein type needed
Recommended method(s)
Cell lysate or tissue homogenate
Intact proteins and protein complexes
Dynabeads for immunoprecipitation
His-tagged recombinants
Intact, recombinant proteins
His-tag purification with affinity resins
Complex protein mixtures
Intact antibodies, IgG or Ig fragments
Antibody purification with protein A/G
Fusion proteins
Intact, untagged proteins
Protein tag removal with proteases
Bioproduction
Clean, intact, sterile large- or pilot-scale

protein for industrial applications
Analytical and production chromatography
Whole cells
Intact protein, separated by isoelectric charge
Protein sample fractionation
Whole cells
Intact organelle proteins
Organelle isolation kits
Whole cells
Intact, native protein
Cell lysis buffers and protein extraction kits
Dynabeads for immunoprecipitation
Type of sample
Dynabeads Protein A
or Dynabeads Protein G
Efficient and reliable immunoprecipitation
Reduce protocol time to ~30 minutes

Isolate intact proteins and protein complexes
Minimize background caused by nonspecific binding
Kits supplied with premade buffers for convenience and
Add sample with antibody
increased consistency
Y
Y
For efficient immunoprecipitation, you need only a Dynabeads

Immunoprecipitation Kit, your specific antibody of choice, and a
DynaMag magnet. The protocol takes place in a single tube with
just a few handling steps.
Elute
Add sample with target protein
Y
Y
Y
Y
Y
Isolate pure
antibodies
Elute
Y Y
1. Bind antibodies to the Dynabeads (10 min)

2. Wash once using a magnet (2 min)
3. Mix Dynabeads and sample (e.g., lysate) for immunocapture
(10 min)
4. Wash 3 times using a magnet (5 min)
5. Elute the antigenantibody complex in a small volume
(2 min), or resuspend directly in SDS/LDS sample buffer,
followed by heating
6. Run on a gel
Mild elution for isolation

of native protein
Denaturing elution for isolation

of denatured target protein
Immunoprecipitation in only 30 minutes. Dynabeads precoupled with protein A or

protein G act as a suspendable solid support that can be fixed by the use of a magnet.
This allows for simple and efficient antibody capture, followed by immunoprecipitation of your pure target peptides, proteins, protein complexes, or other antigens.
129
Protein analysis
Gentle, rapid handling for better results
Immunoprecipitation with Dynabeads is fast and gentle, causing minimal physical stress to target proteins.
Rapid kinetics and short incubation times reduce the protocol time typically to only 30 minutes. Dynabeads
permit the isolation of labile complexes that might otherwise dissociate or be damaged by proteases during
long incubations. Native protein conformation and intact, large protein complexes are preserved. Gentle
magnetic handling allows you to work with concentrated protein solutions throughout the procedure. You do
not need to worry about losing material from spun-down resins or excess surface during pipetting. Washing
and elution are very efficient and help ensure maximal sensitivity, minimal loss of target protein, and no
background caused by nonspecific binding.
Easy, flexible, and automatable
The protocol can be scaled to match your specific purpose and sample volume requirements. An important
advantage of short incubation times and the low nonspecific binding characteristics of Dynabeads products
is that there is no need for time-consuming preclearing or dilution.

The table below outlines our most popular immunoprecipitation products. If you go to lifetechnologies.com/
immunoprecipitation, you can find information about our entire line of immunoprecipitation products and
youll also have access to:
Product selection guides for immunoprecipitation (IP) and co-IP

Yield, reproducibility, and binding specificity data for Dynabeads products
Protocols for IP crosslinking and other applications
Information to help you choose the right assay format and magnet
References that cite the use of Dynabeads for products IP and protein isolation
Dynabeads products for immunoprecipitation. Find the immunoprecipitation product you need using our interactive selec
tion guide at youtube.com/immunoprecipitation.
130
Dynabeads product
Bead characteristics
Binding properties
Type of ligand
Coupling
Protein A
(for IP)
Surface:protein A
(Cat. No. 100-06D)
Noncovalent antibody binding
Manyantibodies
>10 min
room temp
Protein G
(for IP)
Surface:protein G
(Cat. No. 100-07D)
Noncovalent antibody binding
Many antibodies
1040 min
room temp
Co-IP kit for protein

complexes
Surface: epoxy
(Cat. No. 143-21D)
Covalent antibody binding

(antibodywill notelute off beads)
Any antibody
15 min setup
(overnight reaction)
His-tag isolation and

pulldown
Surface: cobalt NTA

chemistry
(Cat. No. 101-03D)
Ionic interaction, very low

background
Binds histidine-tagged
proteins
5 min
room temp
Antibody Coupling Kit
Surface: epoxy
(Cat. No. 143-11D)
Covalent binding of antibodies

andother desired proteins
Binds any antibody

andprotein ligand
15 min setup
(overnight reaction)
Protein analysis
His-tag purification with affinity resins
Life Technologies offers affinity resins and purification kits for use with 6xHis-tagged proteins. Purification of histidine-tagged recombinant proteins can be performed with precharged Ni resin (Ni-NTA agarose
beads) or with an iminodiacetic acid ligand (ProBond Nickel Chelating Resin). To select your product, visit
lifetechnologies.com/histagpurification.
Antibody purification with protein A/G
Life Technologies offers a variety of affinity resins for antibody purification. IgG purification, as well as fractionation of IgG fragments, can be achieved using Protein A Agarose, Protein G Agarose, or Streptavidin
Agarose. To learn more or select the product you need, visit lifetechnologies.com/antibodypurification.
Protein tag removal with proteases
Life Technologies offers a wide selection of proteases for efficient removal of affinity tags and cleavage of
fusion proteins. Recombinant proteins that enterokinase, or SUMO recognition sequences provide the option
for protein tag removal with highly specific and active cleavage enzymes.
AcTEV Protease
EKMax Enterokinase
SUMO protease
Cat. Nos.
12575015, 12575023
E18001, E18002
12588018
Enzyme
Tobacco Etch Virus (TEV) with

improved specificity, activity, and
stability than native TEV protease;
contains 6xHis for easy removal
Recombinant bovine enterokinase*
Recombinant fragment of ULP1 from

S. cerevisiae that contains 6xHis for
easy removal
Sequence recognized
Glu-Asn-Leu-Tyr-Phe-Gln-Gly
Asp-Asp-Asp-Asp-Lys
Tertiary structure of SUMO peptide
Used for
Cleavage of any fusion protein

containing the recognition sequence
Cleavage of any fusion protein

containing the recognition sequence
Removal of SUMO fusion tags
Protease
*Use EK-Away Resin (Cat. No. R18001) to remove EKMax Enterokinase (or other enterokinases) from your fusion protein following digestion.
For more information on these products or to place an order, visit lifetechnologies.com/proteases.
Analytical and production chromatography
Weve developed chromatography resins and systems that accommodate protein production at every scale
from pilot projects up to process-scale separations. Choose one of our high-resolution, high-capacity analytical and process resins or one of our specialty resins, designed for specific applications such as large and
small protein biomolecules, viruses, and antibodies.
Product
Application
Particle size
Online resource
POROS Perfusion
Chromatography Resin
High performance, high throughput for

process-scale bioseparations
50 m
lifetechnologies.com/porosproduction
POROS Analytical
Chromatography
Separation of biomolecules for both analytical

and lab-scale preparation
10 and 20 m
lifetechnologies.com/porosanalytical
GoPure Pre-packed
Chromatography Columns
Biopharmaceutical manufacturers in earlyphase process development
20 and 50 m
lifetechnologies.com/gopure
Resins for microbial

expression systems
Microbial protein purification
50 m
lifetechnologies.com/microbialresins
Resins for viral vaccine

production
Purification of viruses or virus-like particles

for vaccine production
50 m
lifetechnologies.com/virusresins
Resins for monoclonal antibody

production
Pilot- or production-scale antibody

purification
10, 20, or 50 m
lifetechnologies.com/antibodyresins
For more information, visit lifetechnologies.com/proteinchromatography.
131
Protein analysis
Protein sample fractionation
The ZOOM IEF Fractionator reduces highly complex protein samples into fractions,
based upon isoelectric point, for analysis by two-dimensional gel electrophoresis
(2DE), one-dimensional gel electrophoresis (1DE), or two-dimensional liquid chromatography/mass spectrometry (2D LC/MS). The ZOOM IEF Fractionator offers a range
of rapid separation (fractionation) options and:
Allows loading of high amounts of protein for downstream applications

Enriches low-abundance proteins and increase the dynamic range of detection
Reduces precipitation/aggregation artifacts associated with highprotein load
samples
ZOOM IEF Fractionator
To learn more about the Zoom Fractionator and reagents, visit lifetechnologies.com/
proteinfractionation.
Organelle isolation kits
To reduce sample complexity prior to analysis, isolate organelles with magnetic

separation technology. Dynabeads can been used to isolate a number of different
organelles and organelle-specific proteins. The magnetic separation technology
enables specific isolations of true fractions in small volumes, and can replace timeconsuming and labor intensive density gradient centrifugations.
Rapid, high-purity isolations from crude fractions or fractions of common

densities
Morphologically intact target organelles can be fixed for EM studies or solubilized

directly on the bead surface
Immunomagnetically isolated organelles can be assayed by all conventional

techniques
Easy protocol yields high-purity functional organelles of interest

Examples of organelles or organelle fractions that have been isolated with Dynabeads
products include mitochondria, peroxisomes, kidney glomeruli, and endocytic and
secretory compartments.
Need
Application requirement
Recommended product
Advantage
Isolation of fragile
organelles or organelle
fractions
Mouse primary antibody
Dynabeads M-280 Sheep AntiMouse IgG

(Cat. No. 112-01D)
Direct or indirect isolation of organelles

made more efficient with two layers of
antibodies
Rabbit primary antibody
Dynabeads M-280 Sheep AntiRabbit IgG

(Cat. Nos. 112-03D, 112-04D)
Direct or indirect isolation of organelles

made more efficient with two layers of
antibodies
Non-mouse/non-rabbit
primary antibody
Dynabeads Protein G (Cat. No.100-03D)

Dynabeads Protein A (Cat. No. 100-01D)
Efficient direct or indirect isolation of

organelles
Isolation of large organelle fractions or isolation

from a viscous sample
Antibodies made to
specific species
Dynabeads M-450 with a secondary antibody against the corresponding primary

antibody species (Cat. No. 140-13)
More efficient pull to the magnet in

viscous samples
Isolation of organelles
using a non-antibody
ligand
High yield
Dynabeads M-280 Tosylactivated

(Cat. No. 142-03)
Easy coupling of a molecule onto these

hydrophobic beads for efficient organelle
isolation
High purity
Dynabeads M-270 Epoxy (Cat. No. 143-01)
Coupling of a molecule onto these

hydrophillic beads for very high-purity
organelle isolation
To learn more about organelle isolation with Dynabeads or to find products, visit
lifetechnologies.com/organelleisolation.
132
Protein analysis
Cell lysis buffers and protein extraction kits
Easy-to-use buffers and kits allow rapid and simple protein extraction as well as
protein extraction combined with RNA isolation from cultured cells. The table below
lists some of our top lysis and extraction products.
Best for
Cat. No.
Cell Extraction Buffer*
Sample prep of cells prior to ELISA and western blot
FNN0011 (standard buffer, 100 mL),

FNN0091 (denaturing buffer, 100 mL)
NP-40 Cell Lysis Buffer*
Sample prep of cells prior to Luminex assays, ELISA, western blot;

contains NP-40 detergent
FNN0021 (100 mL)
Tissue Extraction Reagent 1*
Total protein extraction from tissue samples
FNN0071 (100 mL)
PARIS Kit**
Isolating total RNA and total protein from fresh cultured cells;
alternatively, cells can be partitioned into nuclear and cytoplasmic
fractions before protein/RNA fractionation; phenol-free formulation;
useful for enzymatic assays, immunoprecipitation, gel shift assays,
2D electrophoresis, and western blotting
AM1921 (50 purifications)
mirVana PARIS Kit**
Correlating mRNA, miRNA, or siRNA levels with protein levels;

achieves simultaneous extraction of native protein and RNA from
cells or tissues
AM1556 (2040 isolations)
Buffer
* Go to lifetechnologies.com/proteinlysisbuffer for more information.

** Go to lifetechnologies.com/proteinextractionkits for more information.
133
Protein analysis
Qubit 2.0 Fluorometer

Simple, accurate protein quantitation
The Qubit 2.0 Fluorometer together with the Qubit Protein Assay offer an easyto-use method for rapid quantitation of protein samples. The Qubit 2.0 Fluorometer
reads the signal from an optimized Molecular Probes dye that is selective for protein
even in the presence of an equal mass of DNA or RNA.
Qubit fluorometric quantitation is:
Selectivea protein-selective dye in the Qubit Protein Assay has a linear

detection range of 0.255 g of protein (effective for starting sample
concentrations from 12.5 g/mL to 5 mg/mL)
Sensitivesamples with concentrations as low as 12.5 g/mL of protein may be
accurately and reliably quantitated
Simple and intuitivethe Qubit 2.0 Fluorometer offers rapid results and is easy
to use
The Qubit Fluorometric Quantitation System.
Features of the Qubit 2.0 Fluorometer include:
Protein quantitation
Large LCD color touch screen

Automatic data logging and USB port for data management
Easy workflow navigation
Standard curve display after calibration is completion
A simple protocol gives you quantitation data in minutes
The Qubit assays for use with the Qubit 2.0 Fluorometer are all performed using
the same simple mix-and-read protocol. The Qubit Protein Assay Kits (Cat. Nos.
Q33211 and Q33212) provide concentrated assay reagent, 1X buffer, and prediluted
BSA standards. Simply dilute the reagent using the buffer provided, add your sample
(any volume between 1 L and 20 L is acceptable), incubate for 15 minutes, and read
the concentration using the Qubit 2.0 Fluorometer.
Standards
from kit
10 L
10 L
Add Quant-iT working

solution for a total
volume of 200 L
Vortex tubes
for 23 sec
User
samples
120 L
120 L
120 L
Add Quant-iT working

solution for a total
volume of 200 L
Incubate at room
temperature for
2 min (DNA, RNA)
or 15 min (protein)
Read tubes in
Qubit Fluorometer
Workflow for the Qubit assay using the Qubit 2.0 Fluorometer.
134
Protein analysis
The Qubit Protein Assay exhibits very low
protein-to-protein variability
Compared to other protein assays, the Qubit Protein Assay exhibits very low proteinto-protein variability (see figure). For the most accurate results when quantitating
protein with the Qubit Protein Assay, use detergent-free protein samples. To see
which contaminants are tolerated by the Qubit Protein Assay, see Table 2 of the
Qubit Protein Assay Kits manual.
You can download the manual at lifetechnologies.com/qubitproteinassay.
Pierce BCA Protein Assay
Protein concentration determined

by assay (mg/mL)
BSA
Lysozyme
Histone
Bovine pituitary extract
0.700
0.600
0.400
0.400
0.300
0.200
0.100
0.000
-0.100
0.1
0.2
0.3
0.4
0.5
0.6
0.800
BSA
Lysozyme
Histone
0.700
0.600
0.400
0.400
0.300
0.200
0.100
0.000
-0.100
0.1
[Protein] (mg/mL)
0.400
0.300
0.200
0.100
0.000
0
0.1
0.2
0.3
0.4
[Protein] (mg/mL)
0.5
0.6

by assay (mg/mL)

by assay (mg/mL)
BSA
Lysozyme
Histone
0.400
-0.100
0.4
0.5
0.6
Pierce Lowry Protein Assay
Bio-Rad Quick Start Bradford Protein Assay
0.600
0.3
[Protein] (mg/mL)
0.800
0.700
0.2
Protein quantitation

by assay (mg/mL)
Qubit Protein Assay

0.800
0.800
BSA
Lysozyme
Histone
Bovine pituitary
extract
0.700
0.600
0.400
0.400
0.300
0.200
0.100
0.000
-0.100
0.1
0.2
0.3
0.4
0.5
0.6
[Protein] (mg/mL)
Protein-to protein variability. Triplicate samples of BSA, lysozyme, histone, and bovine pituitary extract were
assayed at concentrations of 00.5 mg/mL using the Qubit Protein Assay on the Qubit 2.0 Fluorometer, and
various other protein quantitation methods. The variability at 0.3 mg/mL for the Qubit assays was found to be <7%.
135
Protein analysis
Gel electrophoresis
High-performance, high-quality precast gels

and gel casting essentials
If you are doing standard one-dimensional protein separation, we have Novex solutions, and this section will help you select the separation tools that fit your research.
The Novex gel line consists of several different chemistries, well formats, and gel
sizes, so you get the separation you need for accurate downstream results.
To learn more about our basic denaturing protein separation,
visit lifetechnologies.com/buyproteingels.
Protein gel electrophoresis
One-dimensional protein separation.

Gel-casting
accessories
Tris-glycine gels
NuPAGE gels
Bolt gels
Application
Pour-your-own essentials:
preassembled empty
cassettes, buffers,
reagents
Precast convenience with

Laemmli chemistry
Straight protein bands,

longest shelf life
Optimized for western blot

performance
Protein resolution
Medium
Medium
High
High
Protein integrity
Medium
Medium
High
High
Shelf life
NA
12 months
8 months (Tris-acetate)
16 months (Bis-Tris)
16 months
Average run time
90 min
90 min
35 min
35 min
Separation range
6500 kDa
6500 kDa
1.5300 kDa (Bis-Tris)

30400 kDa (Tris-acetate)
1.5300 kDa
Gel % available
NA
4%, 6%, 8%, 10%, 12%,

420%, 412%, 14%, 16%,
18%, 816%, 1020%
412%, 8%, 10%, 12%

(Bis-Tris)
38%, 7% (Tris-acetate)
412%, 8%, 10%, 12%
Pack sizes available
NA
5- and 10-packs
2- and 10-packs
10-pack
Gel dimensions
Mini: 8 x 8 cm, 1 or
1.5mm thick
Midi: 8 x 13 cm, 1mm
thick
1.5mm thick
thick
1.5mm thick
thick
8 x 8 cm, 1 mm thick, with

wedge-shaped wells
Maximum sample
volume/well
25 L (1 mm)
37 L (1.5 mm)
25 L (1 mm)
37 L (1.5 mm)
30 L (1 mm)
43 L (1.5 mm)
60 L
Maximum protein load

per band for optimal
resolution*
NA
0.5 g
0.5 g
0.5 g
*On 1.0 mm, 10-well gel, detected by Coomassie staining.
136
Protein analysis
Bolt gels
Bolt Bis-Tris Plus Gels are optimized to deliver superior western blot
performance:
Superior band quality and band volumeattributed to Novex Bis-Tris Plus

chemistry for delivery of sharp, straight bands
Better protein resolution with more protein bands detecteddue to 10%

greater resolving distance and optimized gradient format
Preserved protein integrity with the legendary Novex neutral-pH formulation

that minimizes protein modifications
Best lot-to-lot consistencyour best-in-class manufacturing delivers highly
Bolt Bis-Tris Plus Gels.
reproducible gel performance, with CV of only 2% for Rf resolution values
Bolt gels are designed to deliver optimal resolution, band quality, and sensitivity
for western blot analyses. Bolt gels give well-resolved, flat, straight western blot
bands compared to other commercially available gels that present rounded, poorly
resolved bands. Besides providing more accurate molecular weight assignments
of your proteins, Bolt gels overall provide better-looking, publication-quality
western blots. Unlike traditional Tris-glycine gels, Bolt Bis-Tris Plus gels are
Bis-Tris HCl buffered (pH 6.4) and have an operating pH of about 7.0. This neutral
pH paired with a unique, gentle sample preparation protocol means your western
protein samples always experience mild, nonacidic conditions. This preserves
protein integrity and minimizes protein modifications to help ensure highly sensitive, accurate western blots every time.
For more information and to place your order, visit lifetechnologies.com/bolt.
Western blot band quality from Bolt gels. A Bolt 412% gradient gel was run at 165 V for 35 min. Two
GST fusion proteins as well as GST were spiked into E. coli lysate (0.6 mg per gel lane) to create series of
decreasing, increasing, or constant amounts of these proteins across the gel. The spiked amounts of the two
fusion proteins ranged from 40 ng to 0.2 ng per gel lane. A WesternBreeze Chemiluminescence Detection
Kit was used with an anti-GST antibody. The image was acquired using an LAS-1000 (FujiFilm) system.
137
Protein analysis
Wedge well for loading up to 2x more protein sample
Bolt gels have new wedge-shaped wells for loading up to twice as much protein
sample in every well.
The new wedge well:
Allows loading of dilute samples
Minimizes sample spillover and cross-well contamination
Enables easy gel loading with standard pipette tips
Delivers better detection sensitivity for low-abundance proteins
Gives better stacking of high molecular weight proteins to give better resolution
Enables greater success with low-affinity western antibodies
Wedge wells on the Bolt Bis-Tris Plus Gel.
NuPAGE gels
The NuPAGE precast gel system is a revolutionary high-performance polyacrylamide

gel system.
Superior protein band resolution and stability

Faster sample run times
Longer product shelf life
Gels are available in both Bis-Tris and Tris-acetate formulations and in a variety of
acrylamide percentages (see table). The unique NuPAGE gel formulations minimize
the smiles and poor resolution seen with other gel types. The neutral-pH gel chemistry and buffer system avoid chemical modifications to the samples that occur in
alkaline environments. In addition, proteins transfer from NuPAGE gels more efficiently, resulting in higher detection signals during western analysis.
Novex NuPAGE 412% Bis-Tris Midi Gel.
For more information on our NuPAGE products and to place an order,

visit lifetechnologies.com/nupage.
Two NuPAGE types to choose from.
138
Gel type
Percentages available
Separation range
Shelf life
Average run time
Applications
Bis-Tris
8%, 10%, 12%, 412%
1.5300 kDa
16 months*
3550 min
Separation of small- to medium-sized

proteins; simple sample reduction; protein
sequencing, mass spectrometry, techniques
where protein integrity is crucial
Tris-acetate
7%, 38%
30400 kDa
8 months*
60 min
Excellent separation of large molecular

weight proteins
Protein analysis
Convenient electrophoresis
Intuitive electrophoresis tank
The Bolt Mini Gel Tank (Cat. No. B4477599) is designed for more intuitive use and convenience than
traditional electrophoresis tanks:
Easy sample loadingwith the new front well configuration

Less running buffer requiredwith two separate gel chambers, you only need to load sufficient buffer
for each gel
Simultaneous visualization of both gelswith streamlined, side-by-side tank configuration

Simplified monitoring of prestained markerswith new white tank stand
Bolt Mini Gel Tank.
Gel tanks and buffers
All Novex gels are available in the 8 x 8 cm mini gel format and a larger 8 x 13 cm midi
size for higher throughput. Run up to two mini gels at a time in the XCell SureLock
Mini-Cell (A, below). Use the XCell4 SureLock Midi-Cell (B, below) to simultaneously
run up to four midi gels. Each midi gel can run up to 26 samples, allowing electrophoresis of over 100 samples at a time. With a Midi Gel Adapter (Cat. No. WA0999), Novex
Midi Gels can be run in a Bio-Rad Criterion Cell.
For more info on these and other products for running Novex gels and to place an
order, visit lifetechnologies.com/electrophoresisaccessories.
The XCell SureLock protein electrophoresis cells. (A) XCell SureLock Mini-Cell (Cat. No. EI0001) for running
one or two NuPAGE or Novex mini gels. (B) XCell4 SureLock Midi-Cell (Cat. No. WR0100) for running up to
four NuPAGE or Novex midi gels.
139
Protein analysis
Additional gels from Novex brands
The Novex family of products offers a variety of options for protein separation,
including gels for isoelectric focusing, resolution of undenatured proteins, low molecular weight proteins and even 2 dimension electrophoresis. Please review the selection table below to find the gel most suited for your research needs.
For more information on our comprehensive line of specialty gels for protein separation
and to place an order, visit lifetechnologies.com/specialtygels.
Specialty protein gels
Quickly analyze
4896 proteins
Analyze posttranslational protein

modifications
Excellent
resolution for
low MW
Optimized
resolution of
active proteases
Superior
resolution of
native proteins
Convenient and
simplified 2D
electrophoresis
Product
E-PAGE gels
Novex IEF gels
Novex Tricine
gels
Novex zymogram
gels
NativePAGE gels
ZOOM 2D
Electrophoresis
System
Applications
Compatible with
high-throughput
systems
Separate native
proteins and study
modifications
Separate low-MW
proteins and
peptides
Separate
proteases based
on size and
activity
Determine size
and purity of
native proteins
Fractionate
proteins before
electrophoresis
and MS
Separation
Range
10200 kDa
pI 3.56.8
220 kDa
10220 kDa
1510,000 kDa
pH 312
Max sample
volume/well
20 L
(E-PAGE 48)
20 L
25 L
20 L
25 L
140 L (diluted)
5%
10%, 16%, 1020%
10% (with gelatin)
312%, 416%
pH 310 NL,
310L, 47, 610,
912 (broad
range)
pI 3.58.5
15 L
(E-PAGE 96)
Gel % available
8% (E-PAGE 48)
6% (E-PAGE 96)
Average run
time
14 min
12% and 416%

(with casein)
2.5 hrs
90 min
90 min
pH 4.55.5,
5.36.3, 6.17.1
(narrow range)
90 min
30 min
(1st dimension)
40 min
(2nd dimension)
140
Shelf life
6 months
2 months
12 months
2 months
6 months
12 months
Gel dimensions
13.5 x 10.8 cm,

3.7mm thick
8 x 8 cm, 1.0mm
thick
8 x 8 cm, 1.0mm
thick
8 x 8 cm, 1.0mm
thick
8 x 8 cm, 1.0mm
thick
7 cm strips,
0.5cm thick
Protein analysis
Accurate molecular weight estimations

Sharp, accurate, prestained and unstained standards
Protein separation is often accompanied by band identification. A variety of Novex
molecular weight (MW) protein standards are available to help you make the most
accurate size estimations possible.
Sharpest bandsclearer estimation of MW compared to thick bands

Broadest MW rangeestimation over a larger range (3.5260 kDa)
Ladder-like separation of bandsevenly spaced pattern across the readable range
for easy identification and comparison
To learn more about protein molecular weight standards and to place an order, go to
lifetechnologies.com/proteinstandards.
SeeBlue
Pre-stained Standard
(Cat. No. LC5625)
BenchMark
HiMark Pre-stained
Pre-stained Protein
Protein Standard
Ladder (Cat. No. 10748010) (Cat. No. LC5699)
SeeBlue Plus2
Pre-stained Standard
(Cat. No. LC5925)
Sharp Pre-stained
Protein Standard
(Cat. No. LC5800)
Application
Basic, routine MW
MW marker for
Tris-glycine gels only
Ideal for large

proteins
Multiple color bands,

easy estimation
Easiest to interpret
MW; best size
estimation
Number of bands
and colors
9 bands
1 color
10 bands
2 colors
9 bands
2 colors
10 bands
3 colors
12 bands
12 colors
Band sharpness
Low
Low
Low
Medium
High
MW range
4250 kDa
10190 kDa
30460 kDa
4250 kDa
3.5260 kDa
Gel compatibility
NuPAGE Bis-Tris;
Tricine, Tris-glycine
Tris-glycine only
Tris-acetate, Trisglycine
NuPAGE Bis-Tris;
Tricine, Tris-acetate,
Tris-glycine
NuPAGE Bis-Tris;
Tricine, Tris-glycine
Novex prestained standards. Find this interactive table online at lifetechnologies.com/prestainedproteinstandards.
Unstained protein standards. Find this interactive table online at lifetechnologies.com/unstainedproteinstandards.
Application
Mark12 Unstained
Standard
(Cat. No. LC5677)
BenchMark Protein
Ladder
(Cat. No. 10747012)
Sharp Unstained
Protein Standard
(Cat. No. LC5801)
HiMark Unstained
Protein Standard
(Cat. No. LC5688)
NativeMark Unstained
Protein Standard
(Cat. No. LC0725)
Routine MW
estimation in
denaturing gel
electrophoresis
MW estimation and
quick orientation
MW estimation and
band identification
MW estimation for
large proteins
MW estimation in native
gel electrophoresis
Number of bands
12
15
12
MW range
2.5200 kDa
10220 kDa
3.5260 kDa
40500 kDa
201,200 kDa
Orientation band(s) No
Yes
Yes
No
No
Number of
0
orientation band(s)
2
(20 and 50 kDa bands
more prominent)
3 (20, 50, and

110 kDa bands more
prominent)
Gel compatibility
NuPAGE Bis-Tris;
SDS-PAGE Tricine,
NuPAGE Bis-Tris;
SDS-PAGE Tricine,
NuPAGE Bis-Tris;
SDS-PAGE Tricine,
Tris-acetate
Tris-glycine,
NativePAGE Bis-Tris,
Tris-acetate
Band sharpness
Low
Medium
High
Low
Low
141
Protein analysis
Sensitive stains for total detection

Life Technologies offers a number of staining reagents for total protein and posttranslationally modified proteins, including
Coomassie G-250 stain, silver stain, and a suite of compatible and highly sensitive fluorescent stains that allow direct quantitation of differential phosphorylation and glycosylation patterns as well as total-protein expression from a single sample on the same
gel. Use the table below to find the stain thats right for your application.
Overview of Novex protein stains. For more information on protein gel stains, visit lifetechnologies.com/proteinstains.
Product
SimplyBlue
SafeStain
SilverQuest Silver
Staining Kit
SYPRO Ruby
Protein Gel Stain
Coomassie Fluor Orange

Protein Gel Stain
Cat. No.
LC6060, LC6065
LC6070
S12001, S12000, S21900
C33250,
Type of stain
Colorimetric; ready-to-use
Colorimetric;
colloidal Coomassie G-250 ready-to-use silver
Fluorescent; ready-to-use
Fluorescent; ready-to-use
Instrumentation
required
No
No
UV transilluminator, bluelight box, or laser scanner
UV transilluminator, bluelight box, or laser scanner
Sensitivity
>7 ng BSA
Subnanogram
Subnanogram
48 ng
Background
Low
Low
Low
Low
Linear dynamic range
770 ng
110 ng
0.251,000 ng
41,000 ng
Multiplex capability
No
No
Yes
No
For densitometry
Yes
Not recommended
Integration of fluorescent
signal
Integration of fluorescent
signal
MS compatibility
Yes
Yes
Yes
Yes
Staining membranes
PVDF
No
Use SYPRO Ruby Protein

Blot Stain
No
Total staining/
destaining time
3 hr; 12 min with microwave protocol
90 min; <60 min with

microwave protocol
90 min (mini gels with new

protocol); overnight for
large-format gels
3060 min
Protocol summary
Rinse, stain, water-based

destain
10 solution changes
Fix, stain, and wash
Fix and stain in one step
SYPRO Ruby protein gel stain for ultrasensitive total

protein detection
Simple staining procedure
Linear quantitation range of 3 orders of magnitude
Linear compatibility with mass spectrometry and microsequencing
SYPRO Ruby Protein Gel Stain is a highly sensitive fluorescent stain for proteins in
1D or 2D gels (see figure). The fast microwave mini gel protocol yields optimal results,
typically in only 90 minutes. For convenience, SYPRO Ruby stain is provided ready to
use and stores at room temperature.
Learn more at lifetechnologies.com/proteinstains.
Sensitive fluorescent staining with SYPRO Ruby
Protein Gel Stain.
142
Protein analysis
Efficient protein transfer for

successful western blotting
iBlot 7-Minute Blotting System
Complete transfer in 7 minutes or less
High blotting efficiency and evenness
Increased blotting reliability and reproducibility
The iBlot 7-Minute Blotting System is designed to work with

multiple polyacrylamide gel sizes, including the midi size, fitting
midi gels (8 x 13 cm) and E-PAGE 48 and 96 gels, and mini gels
(8 x 8 cm).
1 2 3 4 5 6 7 8 9
10 11
12
1 2 3 4 5 6 7 8 9 10 11 12
The iBlot 7-Minute Blotting System (Cat. No. IB1001)

is designed to efficiently and reliably blots proteins from
polyacrylamide gels in 7 minutes or less, without the need for
buffers or an external power supply. A self-contained unit, the
iBlot device uses disposable blotting stacks with integrated
nitrocellulose or PVDF transfer membranes, offering the
convenience of a bufferless, plug-and-play system. In addition
to ease and convenience, the iBlot system offers highefficiency protein transfers (see figure) for high downstream
detection sensitivitiesin many cases, higher than conventional
methods. In the end, you can achieve more accurate detection
using less sample.
The iBlot 7-Minute Blotting System.
High transfer efficiencies achieved using the iBlot 7-Minute Blotting System. (A)
iBlot dry transfer to nitrocellulose and (B) semi-dry transfer to nitrocellulose of
NuPAGE 12% Bis-Tris Mini Gels. Lanes 16: 0.0625 g, 0.125 g, 0.25 g, 0.5 g,
1.0 g, and 2.0 g of SW480 human colon cancer cell lysate (Alexis) (control); lanes
7 and 12: 5 L SeeBlue Plus2 Pre-stained Protein Standard; lanes 811: 0.5 L,
1.0 L, 2.0 L, and 4.0 L of MagicMark XP Western Protein Standard.
Which western transfer system is right for you?
Application
Traditional wet transfer in 12 hr
Semi-dry transfer for high efficiency
Dry transfer in 7 min
Product
XCell II Blot Module

(Cat. No. EI9051)
Novex Semi-Dry Blotter

(Cat. No. SD1000)
iBlot Gel Transfer Device

(Cat. No. IB1001)
Transfer time
60120 min
3060 min
7 min
Capacity of device
12 mini gels
4 mini gels or
2 midi gels
2 mini gels or
1 midi gel
Blotting area
9 x 9 cm
21 x 21 cm
13.5 x 8.5 cm
Requires transfer buffer
Yes (1,000 mL)
Yes (500 mL)
No
Power supply
External
External
Internal
For more information and to place an order for western transfer systems, visit lifetechnologies.com/westerntransfer.
143
Protein analysis
Precut blotting membranes
Simplify blotting setup with precut membrane/filter-paper sandwiches. Life Technologies makes blotting easier by providing a variety of precut, preassembled membrane/
filter-paper sandwiches for mini and midi/E-PAGE gels (see figure). A proteins
properties (i.e., charge, hydrophobicity, etc.) affect its ability to bind to membrane
surfaces. Finding the right membrane may require experimenting with your specific
protein on different membranes.
For more information about our western analysis products, go to
lifetechnologies.com/iblot.
1 2
Western transfer and detection
1 2
Invitrolon PVDF
Other 0.45 m PVDF
High signal achieved on Invitrolon PVDF. A 53 kDa protein containing a c-Myc epitope was transferred to (A)
Invitrolon PVDF and (B) another manufacturers 0.45 m PVDF membrane. Both PVDF membranes were
probed with a 1:500 dilution of mouse anti-Myc antibody, then developed with the WesternBreeze Chemiluminescent Anti-Mouse Kit. Blots shown here are 2 min exposures on X-ray film. Lanes 14: 2 ng, 1 ng, 0.5ng,
and 0.2 ng of protein.
BenchPro 4100 Western

Processing System
Just push run and walk away
Reduces tedious hands-on work and human errors

Minimizes cross-contamination, and no clean-up
Automate without protocol changes
Compatible with all chemiluminescent, chromogenic, and fluorescent immunode-
BenchPro 4100 Western Processing System.
tection reagents and protocols.

The BenchPro 4100 Western Processing System (Cat. No. WP0001) helps consistently and accurately deliver the correct volumes of solutions to the membrane at
precise times. This automation removes the tedious and repetitive liquid handling
steps associated with manual western blot processing. Western blots processed
using the BenchPro 4100 system minimize human processing errors, so you get
greater experimental reproducibility and data consistency.
The BenchPro 4100 Western Processing System is designed to generate consistent,

reproducible results, run after run. It is compatible with all chemiluminescent, chromogenic, and fluorescent immunodetection reagents and protocols (see figure). The
BenchPro 4100 system dramatically reduces the need for protocol optimization and
allows you to perform immunoblotting your way, right away.
For more information, go to lifetechnologies.com/benchpro4100.
144
1
A
Manual processing vs. BenchPro 4100 processing of

western blots. (A) Manual processing. (B) BenchPro
4100 system processing. Lane1: 8 L of a 1:10 dilution
of MagicMark XP Standard; lane 2: 50 ng BSA; lane 3:
25 ng BSA; lane 4: 10 ng BSA. Proteins were detected
using rabbit anti-BSA antibody and WesternBreeze
Chemiluminescent KitAnti-Rabbit. Detection
substrate was added to both membranes, and the
blots were imaged at the same time.
Protein analysis
Novex western blot

detection kits
Life Technologies offers a broad range of reagents and kits for western blot detection.
Whether you are using chromogenic, fluorescent, or chemiluminescence detection
systems, we have the solutions you need.
iBlot Western Detection Kits
Fastcomplete western detection, typically in less than 25 minutes when used

with the iBlot 7-Minute Blotting System
Flexibleworks with mini and midi gels
Easy optimizationallows different conditions to be applied to different sections of
the blot
Western transfer and detection
The iBlot Western Detection Kits (see figure) consist of iBlot detection stacks
and iBlot detection reagents. The kits are available with anti-mouse or anti-rabbit
secondary antibodies and are compatible with chemiluminescent and chromogenic
detection. The iBlot Western Detection Kits offer comparable or better sensitivity
than the conventional protocols for the majority of the antibody-antigen pairs, while
offering significant time savings.
Comparison of an iBlot Western Detection Kit to

a WesternBreeze kit. Proteins from an SW480
(human colon adenocarcinoma cell line) lysate were
transferred using the iBlot 7-Minute Blotting System.
A mouse anti-p53 antibody was used as the primary
antibody. Detection was performed with (A) the WesternBreeze Chemiluminescent KitAnti-Mouse or
(B) the iBlot Western Detection, Chemiluminescent
Kit (AntiMouse). The iBlot western detection kit
detected the proteins with sensitivity comparable to
the WesternBreeze kit.
For more information and to place an order,

visit lifetechnologies.com/westerndetection.
Which western blot detection kit is right for you?
Application
Sensitive chemiluminescent
detection
Fluorescence-based kits
utilizing Qdot technology
Sensitive, completekits
Sensitive western
detection in 30minutes
Product recommended
Novex chemiluminescent
substrates
WesternDot 625 Western

Blot Kits
WesternBreeze Western
Blot Immunodetection
Kits
iBlot Western Detection

Kits
Protocol time (not

including transfer)
~3 hr
~3 hr
~3 hr
< 30 min
Sensitivity
Picogram
<1 ng
Femtogram to picogram
Femtogram to picogram
Detection method
Chemiluminescent
Qdot fluorescent
technology
Chemiluminescent or
chromogenic
Chemiluminescent or
chromogenic
Compatible secondary
antibody conjugates
HRP
Biotin
AP
AP
Blocking and wash

solutions included
No
Yes
Yes
Yes
Secondary antibody
included
No
Yes
Yes
Yes
Instruments required
Film developer/scanner
Blue or UV transilluminator
Film developer/scanner
iBlot Gel Transfer Device
145
Protein analysis
Novex primary antibodies for

western blotting
Designed for specific, high-avidity binding to the target of interest
Superior lot-to-lot consistency and excellent signal-to-noise ratios
Exhibit excellent performance in a variety of immunodetection applications
Life Technologies offers high-quality, highly specific primary antibodies for a broad range of applications and
targets, combining the breadth and quality of Invitrogen, Zymed, Caltag, and Biosource under one
brand: Novex. Our primary antibodies are created in-house or licensed from leading scientists from around
the world, purified using methods designed to retain affinity and ensure stability, and tested in multiple
applications such as western blotting, immunohistochemistry, immunofluorescence, flow cytometry, ELISA,
neutralizing assays, and other functional studies.
Primary antibodies for western
Our selection of antibodies covers a diverse range of research areas, including (but not limited to):
Phospho- and cleavage-sitespecific antibodies

Total (pan) antibodies
Cytokine, chemokine, and growth factor antibodies
-amyloid, tau, and other neuroscience antibodies
CD antibodies
Cell junction (gap and tight) antibodies
Mitochondrial oxidative phosphorylation system antibodies
Epigenetic antibodies
Epitope tag antibodies
Stem cell antibodies
Quickly and easily find exactly the right primary antibody
Life Technologies offers over 2,800 antibodies, and we are adding new ones every day. For the fastest and
easiest way to find the primary antibody you need, use the Primary Antibody Search Tool (lifetechnologies
.com/antibodies). Simply search by target, gene symbol, or gene ID, then filter your results by application,
reactivity, host, conjugate type, or antibody type. You can also search within your results to quickly find the
right antibody.
146
Protein analysis
ABfinity recombinant monoclonal antibodies
Engineered antibodies for consistent results
High performance with proven lot-to-lot consistency

Undergo rigorous validation to help ensure specificity
Enables you to detect low-level targets with less sample
Lot 1
kDa
250 A B
Lot 2
C
Lot 3
C
Lot 4
C
150
100
75
Produced using our proprietary technology, ABfinity recombinant rabbit monoclonal

antibodies are highly specific, high-quality antibodies, superior in consistency and
performance.
SMAD2
50
37
25
ABfinity recombinant antibodies are produced on a large scale by expressing them

in mammalian cells, and appear just like their counterparts isolated from serum or
produced by hybridomas. Intact IgG appears on a nonreducing gel as ~150 kDa band
and upon reduction generates a ~25 kDa light chain and a ~50 kDa heavy chain.
15
Lot-to-lot consistency. HeLa cell extracts were

separated on reducing gels, blotted, and probed with
four lots of SMAD2 ABfinity Antibody. Lanes A, B,
and C were loaded with serial dilutions of the HeLa
extract. The primary antibody was detected using the
WesternBreeze Chemiluminescent Kit Anti-Rabbit.
Find out about the quality and consistency of ABfinity antibodies

at lifetechnologies.com/abfinity.
20
Each ABfinity antibody is manufactured in mammalian cells with antibody heavy
and light chain cDNAs from a functionally screened, immunogen-specific antibody.
This highly reproducible process results in superior lot-to-lot consistency, which helps
save time and money by minimizing or eliminating the need to revalidate performance
on new lots. The figure (right) shows the consistent western blotting results achieved
using independent lots of an ABfinity antibody.
Reliable sensitivity and specificity
The ABfinity technology delivers highly sensitive and specific antibodies that only
react with the target of choice, minimizing detection of the wrong signal due to
nonspecific binding. In addition, high-affinity antibodies allow you to use less antibody
for detection, detect very low-level targets, and conserve your precious samples.
This western blot (right) shows a direct comparison of an ABfinity STAT4 antibody
with the best commercial STAT4 antibodies in western blotting. This comparison
includes antibodies from polyclonal, traditional hybridoma monoclonal, and rabbit
hybridoma monoclonal platforms.
ABfinity
antibody
70185
kDa
150
Polyclonal
antibody
714500
4
Competitor
A
7
Competitor Competitor
B
C
9
10
11
12
13 14
100
75
50
37
25
20
15
Reliable sensitivity and specificity. STAT4 ABfinity

and polyclonal antibodies were used at 2, 1, and
0.5 g/mL. STAT4 antibodies from other vendors
were used at their recommended concentrations. The
primary antibody was detected using the WesternBreeze Chemiluminescent Kit Anti-Rabbit.
Extensive validation and characterization

ABfinity antibodies are validated and characterized by multiple applications. The
figures on the next page are the results of an individual antibody in multiple applications: flow cytometry, immunocytochemistry, immunohistochemistry, and western
blot. This extensive validation process allows confidence in target specificity, without
any need for optimization.
147
Protein analysis
Number of events
250
kDa
150
200
100
150
75
60 kDa
100
50
50
37
100
101
102
103
104
25
Fluorescence
Immunohistochemistry of human esophagus carci

noma tissue labeled with AKT [pS473] ABfinity
Recombinant Rabbit Monoclonal Antibody. Formaldehyde-fixed, paraffin-embedded (FFPE) human
esophagus carcinoma tissue was labeled with rabbit
antiAKT [pS473] (0.5 g/mL). Tissues were pretreated
with EDTA and detected with SuperPicture Polymer
DAB. Image was taken at 20x magnification. Note
nuclear and cytoplasmic staining in tumor cells.
Flow cytometry of Jurkat cells labeled with AKT

[pS473] ABfinity Recombinant Rabbit Monoclonal
Antibody. Jurkat cells were incubated with 50 M
PI3K/AKT signaling pathway inhibitor LY294002 (red
trace) or without (green trace) for 1 hr prior to being
fixed and permeabilized using FIX & PERM reagents.
Cells were then stained with 1 g/test of ABfinity
Recombinant Rabbit Monoclonal Antibody followed by
Alexa Fluor 488 goat antirabbit IgG. The blue trace
represents secondary antibody alone.
Western blot of 3T3 cell lysates labeled with AKT

[pS473] ABfinity Recombinant Rabbit Monoclonal
Antibody. Rabbit antiAKT [pS473] (0.1 g/mL) was
used to label AKT [pS473] in untreated 3T3 lysates
(lane 1) or PDGF-treated 3T3 lysates (lane 2).
Immunocytochemistry of mouse fibroblasts cells labeled with AKT [pS473] ABfinity Recombinant Rabbit
Monoclonal Antibody. Mouse fibroblast cells were treated with (A) or without (B) 10 g/mL insulin and labeled
with rabbit anti-AKT [pS473] (5 g/mL). Signal is knocked down after incubation with the phosphopeptide used
as antibody (C) but not with the nonphosphopeptide (D). Alexa Fluor 488 goat antirabbit IgG at 1:1,000 was
used as secondary antibody. Nuclei are stained with Hoechst (blue).
Recombinant proteins
We offer a wide selection of Novex recombinant proteins for immunology, cancer, and stem cell research.
Affordablechoose the size and price that meet your needs and your budget
Activepure, and tested for biological performance
Relevantvast portfolio of mammalian proteins suited for your research
On our website youll find thousands of recombinant proteins, the majority produced in mammalian systems. Mammalian cell
expression ensures proper folding, glycosylation, gamma carboxylation and other post-translational modifications that make the
protein more physiologically relevant.
Proven performance and comprehensive validation data for our proteins ensures that you have confidence in your research.
Through a partnership with Sino Biologicals, we are now able to offer you our widest selection of membrane, internal and secreted
recombinant proteins, all at extremely competitive prices. Find highly relevant proteins for the study of cancer, immunology, stem
cells and more.
Go to lifetechnologies.com/recombinantproteins to find:
An easily searchable protein product guide
Data demonstrating the purity, biological activity, and functionality of our recombinant proteins and growth factors
The most popular recombinant proteins and their uses
148
Protein analysis
Protein detection
For sensitive, specific detection of intracellular or extracellular proteins, Life Technologies offers a wide range of Novex immunodetection-based products in convenient,
ready-to-use formats. We have developed antibody pair kits and ELISA kits for singleanalyte analysis and Novex multiplex assays for multi-analyte analysis. You can use
our immunoassays to investigate:
Targets
Species
Sample types
Cytokines
Chemokines
Signaling proteins
Receptors
Neurobiology markers
Growth factors
Adhesion molecules
Human
Mouse
Rat
Monkey
Swine
Bovine
Serum
Plasma
Supernatant
Cell lysate
Tissue homogenate
Urine
Cerebrospinal fluid
Protein detection
Go to lifetechnologies.com/immunoassay to find:
Information on all of our antibody pair kits, ELISA kits, and Novex multiplex assay
kits and instruments
Immunoassay Selection Guide that allows you to search for assays based on your
target protein
Data demonstrating assay specificity and sensitivity
Characteristics of our antibody pair kits, ELISA kits, and Luminex Assay Kits. Go to lifetechnologies.com/immunoassay to
find an interactive guide that allows you to search by your protein of interest, then filter by validated application, reactivity, or
assay type to find exactly what you need.
Immunoassay type
Antibody Pair Kit
ELISA Kit
Novex Multiplex Assay Kit
Ready-to-use reagents
No; need overnight coating

process
Yes
Yes
Analytical sensitivity
<10 pg/mL
<10 pg/mL
<10 pg/mL
Dynamic range
5250 pg/mL
5250 pg/mL
52,000 pg/mL
Incubation time
4 hr
2.54 hr
3.5 hr
Multiplexable
No
No
Yes
Number of targets measured/well
150
Readout
HRP-TMB (colorimetric)
R-PE (fluorescent)
Instrumentation needed
Microplate reader
Microplate reader
Luminex instrument (Luminex

100/200, MAGPIX, or
FLEXMAP 3D System)
Instrument read time
2 min
2 min
2060 min
149
Protein analysis
Antibody pair kits

Novex Antibody Pair Kits contain matched, pretitered, and fully optimized capture
(coating) and detection antibodies. These kits enable you to build your own ELISA or any
other assay platform that utilizes a matched antibody pair. For convenience, we also offer
a Buffer Kit for Antibody Pairs that contains premade, easy-to-use buffers and solutions
that are optimized for use with Antibody Pair Kits.
Convenient format for maximum flexibility

Cost savings over ready-to-use ELISA kits
Easy to use with optimized reagents and protocol
Antibody Pair Kit supplies sufficient reagents
for 10 ELISA plates (5 plates for intracellular
targets) and includes:
Assay Buffer Set (Cat. No. CNB0011) supplies

sufficient reagents for 10 ELISA plates and
includes:
Capture antibody
Detector antibody
Recombinant standard
Streptavidin-HRP conjugate
Coating buffer A
Coating buffer B
Assay buffer (5X)
Wash buffer (25X)
Stabilized chromogen (substrate)
Stop solution
Protein detection
Learn more at lifetechnologies.com/buildyourownimmunoassay.
ELISA kits
Novex ELISA kits allow specific, quantitative measurements of proteins including cytokines, chemokines, beta amyloids, and signaling targets. Novex ELISA test kits come
ready to use with a precoated 96-well plate and all necessary reagents. A detailed, easyto-follow protocol with kit-specific performance information is also provided. Just add
sample, run the assay, and typically get quantitative results in half a day.
Specifications of Novex ELISA kits.
150
Type of ELISA
Colorimetric ELISA
Ultrasensitive ELISA
ChemiELISA
Analytical sensitivity
<10 pg/mL
<1 pg/mL
<1 pg/mL
Dynamic range
5250 pg/mL
0.520 pg/mL
0.52,000 pg/mL
Incubation time
4 hr
45 hr
3.5 hr
Readout
AP-CSPD (chemiluminescent)
Instrumentation needed
Microplate reader
Microplate reader
Microplate reader with

chemiluminescence capability
Protein analysis
We research each target protein and calibrate our ELISA kits to provide physiologically relevant sensitivity. In
addition, kits are validated using common sample types including serum, plasma, supernatant, and cell lysates
for signaling or phosphorylation proteins. Our ELISA kits are manufactured to help ensure excellent quality and
reproducibility. Kits must meet quality-controlled specifications for sensitivity, dynamic range, precision, specificity, recovery, and lot-to-lot consistency in order to leave our facilities. Our stringent quality specifications require
that each new lot of ELISA kits exhibits minimal variation compared to previous lots.
Rigorous assay validation of Novex ELISA kits helps ensure consistent, reliable results.
Description
What does it mean?
Standard calibration
Calibrated to NIBSC, if available
Widely used for accurate quantitation and consistent standard of reference
Precision
Inter-assay CV <10%, intra-assay CV <10%
Consistent results each time
Sensitivity
Relevant levels of protein
Enables detection of low levels of proteins
Specificity
Cross-reactivity tests are performed with similar

analytes
Helps to ensure accurate measurement of protein of

interest
Recovery
Buffers are optimized to minimize matrix effects
Helps to ensure accurate quantitation in serum and plasma
In-house controls are tested to measure within set

ranges
Helps to ensure consistent results with new lots
Linearity of dilution
Linear results over the quantitative range of the assay
Serial dilution of samples quantitate accurately
Parallelism
Recombinant protein standards mimic native proteins
Samples can be measured with confidence
Protein detection
Specification
Sensitive, accurate, and consistent performance

Validated on serum, plasma, tissue culture supernatant or lysate
Ready-to-use, convenient assay in only half a day
Learn more about our standard, ultrasensitive, and chemiluminescent ELISA kits at lifetechnologies.com/elisakits.
Chemiluminescent ELISA kits
High sensitivityallows measurement of proteins expressed at low levels in serum and plasma
Large dynamic rangebroad assay range minimizes guesswork of sample dilutions
Consistent resultsimproved precision at low end of quantitative range
10,000
Chemiluminescent ELISA
Ultrasensitive ELISA
Standard ELISA
Signal to noise
1,000
100
10
0.1
10
100
1,000
Concentration (pg/mL)
10,000
Standard curve comparison of Hu IL-1B Chemiluminescent ELISA

Kit (Cat. No. KHC0019), Hu IL-1B Ultrasensitive ELISA Kit (Cat. No.
KHC0014), and the standard Hu IL-1B ELISA Kit (Cat. No. KHC0011).
The chemiluminescent kit has a greater dynamic range than other
formats. In addition, the sensitivity levels of the kit is <1 pg/mL, almost
10-fold greater than standard ELISA kits.
To see specific kits, go to lifetechnologies.com/chemielisas.
151
Protein analysis
Neuro ELISA kits
ELISA kits for accurate A (-amyloid), tau, and -synuclein protein quantitation to
assist Alzheimers disease researchers
Easy-to-run sandwich ELISA

Precoated, breakable 96-well plate
Consistent, accurate, and sensitive measurements
2.4
2.0
OD
1.6
1.2
0.8
Learn more about our neuro ELISA kits at lifetechnologies.com/neuroelisas.
0.4
0
Serial dilution of human CSF containing A1-42 demonstrates linearity over the
range of the assay (Cat. No. KHB3441). Linear regression analysis of samples
versus the expected concentration yielded a correlation coefficient of 0.99.
Cerebrospinal fluid
Protein detection
Dilution
Measured (pg/mL)
Expected (pg/mL)
600
1,200
1,800
Typical 7-point standard curve for Tau ELISA Kit (Cat.
No. KHB0041). The dynamic range is 312,000 pg/mL,
and the analytical sensitivity <12 pg/mL.
Expected %
1:4
341.0
341.0
1:8
185.4
170.5
108
1:16
94.6
85.3
107
1:32
41.2
42.6
97
phosphoELISA kits
Specificitytwo antibodies directed against analyte provides better specificity than
3.0
OD 450 nm
western blotting
Sensitivitymore sensitive than western blotting
Sample volumerequires less sample compared to western blotting
Quantitationget quantitative data (unlike with western blots)
Medium throughput96-well format, results in 4 hours, no densitometry analysis
needed
4.0
2.0
1.0
Learn more about our phosphoELISA kits at lifetechnologies.com/phosphoelisas.
0.63 1.25
2.5
10
20
40
Protein (g)
Quantitative data from (A) STAT5a [pY694] phospho

ELISA assay (Cat. No. KHO0761) confirms that from
(B) western blotting using a NuPAGE Novex gel.
Assays were performed in parallel.
152
Protein analysis
Move up to multiplexing
Multiplexing allows simultaneous analysis of multiple markers in one sample to measure
cytokine secretion and the associated downstream phosphorylated proteins to help
give you the complete story both inside and outside the cell. Life Technologies offers
a complete solution for the Luminex xMAP technology including polystyrene and
magnetic assay kits, reagents, xPONENT systems software, technical support and the
entire Luminex portfolio of instruments including the Luminex 200, FLEXMAP 3D,
and MAGPIX platforms, covering the full range of multiplexing capabilities.
We have a wealth of information for you online at lifetechnologies.com/luminex. There
youll find:
Information about how Novex multiplex assays work
Data on the reliability, calibration standards, and specificity of the assays
Help finding the appropriate Novex multiplex assays through our interactive Immunoassay Selection Guide
Lists of assays grouped by species and target
Our Custom Order Tool for building and ordering your custom Novex multiplex assay
Novex multiplex assay kits offer superior accuracy

Protein detection
100%
Recovery
75%
50%
25%
FG
F
-b
a
1, sic
IL
G- 1b
CS
IL F
-1
IL 0
-1
3
IL
IL 6
Eo -12
ta
xi
IL n
M -17
IP
GM -1
-C a
M SF
IP
M 1b
CP
IL 1
-1
VE 5
G
IF F
N
IL -g
-1
R
TN A
Fa
IL
-2
IL
IP 7
-1
0
IL
-4
IL
-8
0%
Novex assay
Supplier 3
Supplier 2
100%
79%
66%
Recovery
75%
50%
17%
25%
0%
Novex assay
Supplier 3
Supplier 2
To evaluate the Novex multiplex assays, samples of 23 human protein markers were spiked into a sample of human
serum, and the sample was processed using the manufacturers instructions for the Novex multiplex assay kit
and using similar kits from two other suppliers. The multiplex sample was quantified on the MAGPIX system, and
percent recovery was calculated (A) for the individual markers and (B) as an average for the entire group.
153
Protein analysis
Novex multiplex assay kits deliver excellent intra-assay precision
A
100%
8%
7%
7%
75%
6%
CV
CV
5%
50%
4%
3%
25%
3%
3%
2%
1%
0%
FG
Fba
1, sic
IL
G- 1b
CS
IL F
-1
IL 0
-1
3
IL
-6
IL
Eo -12
ta
xi
IL n
M -17
IP
GM -1
-C a
M SF
IP
M 1b
CP
IL 1
-1
VE 5
G
IF F
N
IL -g
-1
R
TN A
Fa
IL
-2
IL
IP 7
-1
0
IL
-4
IL
-8
0%
Novex assay
Supplier 3
Novex assay
Supplier 2
Supplier 3
Supplier 2
To evaluate the Novex multiplex assays, samples of 23 human protein markers were spiked into a sample of human serum, and the sample was processed using the
manufacturers instructions for the Novex multiplex assay kit and using a similar kits from two other suppliers. The multiplex sample was quantified on the MAGPIX
system, and intra-assay precision (CV) was calculated (A) for the individual markers and (B) as an average for the entire group.
Protein detection
Learn more at lifetechnologies.com/luminex.
Novex multiplex magnetic assays
Novex multiplex assays, based on Luminex xMAP (multi-analyte profiling) technology, provide a versatile platform that gives users more flexibility and greater array
options for single or multiple analytes. The new line of magnetic multiplex assays was
carefully designed and tested to help ensure that sensitivity, range, and correlation are
maximized, using the same components as our polystyrene bead Novex multiplex
assays. This means that the magnetic and polystyrene assays perform with comperable quality and consistency.
Advantages of Novex magnetic multiplex assay kits:
Easier wash steps with the handheld 96-well magnetic separator

Enables automation of wash steps
Eliminates the hassle of vacuum manifold washes and clogging of filter plates
Learn more at lifetechnologies.com/luminexassays.
154
Protein analysis
Luminex 200
MAGPIX
1,000
FLEXMAP 3D
FG ILF- 10
ba
si
IL c
-1
G- b
IL CSF
-1
0
(2
IL IL )
-1 2p 6
RA 40p
N 7
T
Eo ES
ta
xi
n
IL
-1
M 7
IP
GM -1a
-C
M SF
IP
M 1b
CP
IL 1
-1
5
EG
F
IL
-5
HG
F
VE
GF
IF
N
-g
M
DC
ITA
C
M
IL IF
-1
R
TN A
Fa
IL
-2
M
IG
IL
-4
IL
-8
Monkey peripheral blood mononuclear cells (PBMC) were stimulated in vitro with PMA and A2318 for 72 hours. Markers were detected using the Monkey Cytokine
Magnetic 28 Plex Panel (Cat. No. LPC0003M), and the results were analyzed on the Luminex 200, MAGPIX, and FLEXMAP 3D Systems.
Protein detection
Luminex instruments for xMAP technology

Learn more about our instruments for evaluating multiplex Luminex assays at lifetechnologies.com/luminexinstrument.
Overview of the Luminex instrument systems.
Product
MAGPIX System
Luminex 100/200 System
FLEXMAP 3D System
Feature
Affordable, efficient, compact size
Versatile and efficient; the

standard in multiplexing
High throughput, high plex,

automation compatible
Optics
LED/CCD camera
Lasers/APDs/PMTs
Lasers/APDs/PMTs
Hardware
Fluorescence imager
Flow cytometry based
Flow cytometry based
Bead compatibility
Magnetic beads only
Polystyrene and magnetic beads
Polystyrene and magnetic beads
Multiplex capacity
50
100
500
Read time for 96-well plate
~60 min
~40 min
~20 min
Dynamic range
3.5 logs
3.5 logs
4.5 logs
Microtiter plate
96-well
96-well
96-well and 384-well
155
Index
Numerical
2D electrophoresis system . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
Basic Fibroblast Growth Factor (bFGF) . . . . . . . . . . . . . . . . . . .61
7-AAD . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
BenchMark Protein Ladders . . . . . . . . . . . . . . . . . . . . . . . . 141

BenchPro 4100 Western Processing System . . . . . . . . . . . . 144
BG-11 growth media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Index
ABfinity recombinant
Blasticidin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
monoclonal antibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
BODIPY ceramides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
AcTEV Protease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Bolt gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
Actinomycin D . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Brain Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Advanced reduced-serum media . . . . . . . . . . . . . . . . . . . . . . 41
Breast Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
AKT pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Alexa Fluor 488 Antibody Labeling Kit . . . . . . . . . . . . . . . . . 106
Alexa Fluor 488 Goat AntiMouse IgG (H+L) . . . . . . . . . . . . 106
Calcein AM probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Alexa Fluor dyelabeled phalloidins . . . . . . . . . . . . . . . . . . . . 90
Cancer research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Algae engineering kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
Carbenicillin, disodium salt . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
AlgiMatrix 3D cell culture system . . . . . . . . . . . . . . . . . . . . . . 49
CD Hybridoma Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Alkaline Phosphatase Live Stain . . . . . . . . . . . . . . . . . . . . . . . . 66
CD3/CD28 Dynabeads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Amino acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
CD3/CD28/CD137 Dynabeads . . . . . . . . . . . . . . . . . . . . . . . . . 79
AmnioMAX media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Cell counting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Ampicillin sodium salt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Cell culture and cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Analytical and production chromatography . . . . . . . . . . . . . . 131
Cell depletion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Antibiotics and antimycotics . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Cell dissociation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Antibody and immunoassay selection guides . . . . . . . . . . . . . 37
Cell Extraction Buffer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
Antibody Coupling Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
Cell Line Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Antibody Pairs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
Cell lysis buffers and protein extraction kits . . . . . . . . . . . . . 133
Antibody purification with protein A/G . . . . . . . . . . . . . . . . . . 131
Cell structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
Antifoaming agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Cell tracing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Anti-miR Inhibitors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
Cell tracking and neuronal tracing probes . . . . . . . . . . . . . . . .95
Aortic Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Cell viability, proliferation, and health . . . . . . . . . . . . . . . . . . . . 81
APEX Antibody Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . 107
CellEvent Caspase-3/7 Green Detection Reagent . . . . . . . . . 85
Assay Buffer Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
CellLight reagents
Astrocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Actin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
Astrocyte medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
ER . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Attune Acoustic Focusing Cytometer . . . . . . . . . . . . . . . 55, 100
Golgi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Histone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
156
Lysosomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
B-27 Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
MAP4 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
BackDrop Background Suppressor . . . . . . . . . . . . . . . . . . . 111
Mitochondria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
BacMam ion channel targets . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
Nucleus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Bac-N-Blue Baculovirus
Peroxisomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Expression System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
Tubulin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
Bacterial selection antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . 46
CellROX Deep Red Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
Bac-to-Bac Baculovirus Expression System . . . . . . . . . . . . 117
CELLstart CTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
BaculoDirect Baculovirus Expression System . . . . . . . . . . 117
CELLstart CTS Substrate . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
BaculoDirect Linear DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
CellTrace reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83, 95
Baculovirus expression system . . . . . . . . . . . . . . . . . . . . . . . . 117
CellTracker probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Balanced salt solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Cellular Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Index
E
Champion Expression Systems . . . . . . . . . . . . . . . . . . . . . . 119
EKMax Enterokinase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Champion Vector Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
ELF 97 Endogenous Phosphatase

Detection Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
Chemically Defined (CD) Hybridoma Media . . . . . . . . . . . . . . . 42
ELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .150
Chemiluminescent ELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . 151
Embryonic stem cellqualified FBS . . . . . . . . . . . . . . . . . . . . . 44
Chemiluminescent substrates
Endoplasmic reticulum and Golgi apparatus . . . . . . . . . . . . . .93
for western detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145
E-PAGE gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
Chemokines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .47
Epidermal keratinocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Click-iT EdU Cell Proliferation Assays . . . . . . . . . . . . . . . . . . 82
Episomal reprogramming vectors . . . . . . . . . . . . . . . . . . . . . . 64
Click-iT TUNEL Alexa Fluor Imaging Assay . . . . . . . . . . . . . 85
ER-Tracker dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Co-IP kit for protein complexes . . . . . . . . . . . . . . . . . . . . . . . . 130
Essential 8 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Collagenase IV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Ethidium homodimer-1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Concanavalin A Alexa Fluor 594 conjugate . . . . . . . . . . . . . . 93
Eukaryotic selection antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . 46
Coomassie Fluor Orange Protein Gel Stain . . . . . . . . . . . . 142
Expi293 Expression System . . . . . . . . . . . . . . . . . . . . . . 42, 116
Corneal Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Express Five SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Countess Automated Cell Counter . . . . . . . . . . . . . . . . . . . . . 53
Extracellular matrices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Index
Charcoal-stripped FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Cryopreservation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
CSM media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Custom biology services . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33
FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Custom media for primary cells . . . . . . . . . . . . . . . . . . . . . . . . 54
Fixation and permeabilization

for flow cytometry samples . . . . . . . . . . . . . . . . . . . . . . . . . . 105
Custom Novex multiplex assays
for Luminex technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
FLEXMAP 3D System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Custom primary cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
FLoid Cell Imaging Station . . . . . . . . . . . . . . . . . . . . 55, 66, 101
Customer and technical support . . . . . . . . . . . . . . . . . . . . . . . . 37
Flp-In T-REx System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116
Cytogenetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Fluo-4 Direct Calcium Assay Kit . . . . . . . . . . . . . . . . . . . . . . 88
Cytokines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Fluorescence SpectraViewer . . . . . . . . . . . . . . . . . . . . . . . 36, 111
CytoTune-iPS Sendai Reprogramming Kit . . . . . . . . . . . . . . . 64
Fluorescent dextran conjugates . . . . . . . . . . . . . . . . . . . . . . . . 95

Fluorescent microspheres . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
FluoroMyelin Green
Dialyzed FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Fluorescent Myelin Stain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Dispase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
FluxOR Potassium Ion Channel Assay . . . . . . . . . . . . . . . . . 88
Drosophila Expression System (DES) . . . . . . . . . . . . . . . . . . 117
FoamAway Irradiated AOF . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Drosophila S2 Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
FreeStyle Expression Systems . . . . . . . . . . . . . . . . . . . . 42, 116
Dulbeccos Modified Eagle Medium (DMEM) . . . . . . . . . . . . . . 41
FreeStyle MAX Transfection Reagent . . . . . . . . . . . . . . . . . 125
Dynabeads cell isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76

Dynabeads for immunoprecipitation . . . . . . . . . . . . . . . . . . . 129
Dynabeads M-270 Epoxy . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
G-5 Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Dynabeads M-280 Sheep AntiMouse IgG . . . . . . . . . . . . . . 132
Gel-casting accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
Dynabeads M-280 Sheep AntiRabbit IgG . . . . . . . . . . . . . . 132
Geltrex LDEV-Free hESC Qualified . . . . . . . . . . . . . . . . . . . . . 62
Dynabeads M-280 Tosylactivated . . . . . . . . . . . . . . . . . . . . . 132
Geltrex LDEV-Free matrix products . . . . . . . . . . . . . . . . . . . . 49
Dynabeads M-450 with secondary antibody
GeneArt Algae Engineering Kits . . . . . . . . . . . . . . . . . . . . . . 120
against primary antibody . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

Dynal CD34 Progenitor Cell Selection System . . . . . . . . . . . 73
DynaMag magnet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
GeneArt gene synthesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121

GeneArt GeneOptimizer
sequence optimization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
Geneticin (G-418) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
157
Index
Human Microvascular Endothelial Cell,
Gibco Cell Culture Basics . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
adult dermis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Gibco custom media services . . . . . . . . . . . . . . . . . . . . . . . . . 30
Human Microvascular Endothelial Cell,
GlutaMAX media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
neonatal dermis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Index
GoPure Pre-packed
Chromatography Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Human Neural Stem Cells (H9-Derived) . . . . . . . . . . . . . . . . . 71
Graces Insect Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Human Pulmonary Artery Endothelial Cell . . . . . . . . . . . . . . . 56
Growth factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Human Pulmonary Artery Smooth Muscle Cell . . . . . . . . . . . 56

Human Skeletal Myoblasts, large size . . . . . . . . . . . . . . . . . . . 56
Human Skeletal Myoblasts, small size . . . . . . . . . . . . . . . . . . . 56
Heart Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Human Ultimate ORF Clones . . . . . . . . . . . . . . . . . . . . . . . . 122
Hematopoietic stem cell culture . . . . . . . . . . . . . . . . . . . . . . . . 73
Human Umbilical Vein Endothelial Cell . . . . . . . . . . . . . . . . . . 56
Hematopoietic stem cells (HSCs) . . . . . . . . . . . . . . . . . . . . . . . 73
Hybridoma-SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
HEPES buffers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Hygromycin B . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Hibernate media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
High Five cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
High Five Cells (BTI-TN-5B1-4) . . . . . . . . . . . . . . . . . . . . . . . 47
iBlot 7-Minute Blotting System . . . . . . . . . . . . . . . . . . . . . . . 143
HiMark Protein Standards . . . . . . . . . . . . . . . . . . . . . . . . . . 141
iBlot Western Detection Kits . . . . . . . . . . . . . . . . . . . . . . . . . 145
His-tag purification with affinity resins . . . . . . . . . . . . . . . . . . 131
IEF gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
HulaMixer Sample Mixer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
Image-iT Fixation/Permeabilization Kit . . . . . . . . . . . . . . . . 111
Human Aortic Endothelial Cell . . . . . . . . . . . . . . . . . . . . . . . . . 56
Image-iT FX Signal Enhancer . . . . . . . . . . . . . . . . . . . . . . . . 110
Human Aortic Smooth Muscle Cell . . . . . . . . . . . . . . . . . . . . . . 56
Image-iT LIVE Lysosomal and
Human Astrocyte Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Nuclear Labeling Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Human Astrocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Human cell isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
Nuclear Labeling Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
Human Corneal Epithelial Cell . . . . . . . . . . . . . . . . . . . . . . . . . 56
iMATCH tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Human Coronary Artery Smooth Muscle Cell . . . . . . . . . . . . . 56
ImMedia Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Human Dermal Fibroblasts, adult . . . . . . . . . . . . . . . . . . . . . . 56
Immunology research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Human Dermal Fibroblasts, neonatal . . . . . . . . . . . . . . . . . . . 56
Insect cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Human Epidermal Keratinocyte, adult . . . . . . . . . . . . . . . . . . . 56
Insect expression systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
Human Epidermal Keratinocyte,
Instruments and accessories . . . . . . . . . . . . . . . . . . . . . . . . . . 99
adult (animal productfree) . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Invivofectamine 2.0 Reagent . . . . . . . . . . . . . . . . . . . . . . . . . 125
Human Epidermal Keratinocyte, neonatal . . . . . . . . . . . . . . . . 56
Ion Proton Sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
Human Epidermal Keratinocyte,

neonatal (animal productfree) . . . . . . . . . . . . . . . . . . . . . . . . 56
Human Epidermal Keratinocytes, pooled . . . . . . . . . . . . . . . . . 56
Human Hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Human Mammary Epithelial Cell . . . . . . . . . . . . . . . . . . . . . . . 56
Human Melanocyte-adult, lightly pigmented . . . . . . . . . . . . . . 56
Human Melanocyte-neonatal,
darkly pigmented . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Human Melanocyte-neonatal, lightly pigmented . . . . . . . . . . 56
Human Melanocyte-neonatal,
moderately pigmented . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
158
Image-iT LIVE Mitochondrial and
J
JAK/STAT pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
JC-1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
Jump-In Fast Gateway System . . . . . . . . . . . . . . . . . . . . . . 116
Jump-In TI-Gateway System . . . . . . . . . . . . . . . . . . . . . . 116
K
Kanamycin sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
KaryoMAX reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
KnockOut media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60, 61
Index
L
N
Labeled secondary antibodies . . . . . . . . . . . . . . . . . . . . . . . . . 109
N-2 Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Labeling and detection products . . . . . . . . . . . . . . . . . . . . . . . 106
NativeMark Unstained Protein Standard . . . . . . . . . . . . . . .141
Large-scale protein labeling kits . . . . . . . . . . . . . . . . . . . . . . 107
NativePAGE gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140

Natural Mouse Laminin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
NBD C6-ceramide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Lectin HPA Alexa Fluor 488 conjugate . . . . . . . . . . . . . . . . . . 93
Negative isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Lentiviral iPSC reprogramming particles . . . . . . . . . . . . . . . . . 64
Neomycin sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Lipofectamine reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
Neon Transfection System . . . . . . . . . . . . . . . . . . . . . 55, 65, 124
LIVE/DEAD Viability/Cytotoxicity Kits . . . . . . . . . . . . . . . . . . . 81
Neural stem cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Liver Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Neural stem cell differentiation . . . . . . . . . . . . . . . . . . . . . . . . 72
Luminex 100/200 System . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Neural stem cells (NSCs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .71
Luminex technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
Neuro ELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
LysoSensor dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Neurobasal Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48, 71
Lysosomes and peroxisomes . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Neurobiology media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
LysoTracker dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Neurobiology research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Index
LB medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
LC3B Antibody Kit for Autophagy . . . . . . . . . . . . . . . . . . . . . . . 86
Neurodegeneration pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
M
M9 minimal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
MagicMedia Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
MAGPIX System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Mammalian cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Mammalian expression systems . . . . . . . . . . . . . . . . . . . . . . . 115
MAPK pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Mark12 Unstained Standard . . . . . . . . . . . . . . . . . . . . . . . . . 141
MarrowMAX Bone Marrow Medium . . . . . . . . . . . . . . . . . . . . 51
Melanocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Membrane tracers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Mesenchymal stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
Neuronal tracing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
NeuroTrace Nissl stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Novex Semi-Dry Blotter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
NP-40 Cell Lysis Buffer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
NucBlue Fixed Cell Stain
(DAPI special formulation) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
NucBlue Live Cell Stain
(Hoechst 33342 special formulation) . . . . . . . . . . . . . . . . . . . 94
Nucleus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
NuPAGE gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
MesenPRO RS Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
Opti-MEM reduced-serum medium . . . . . . . . . . . . . . . . . . . . 41
Microbial culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Organelle isolation kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
Microvascular endothelial cells . . . . . . . . . . . . . . . . . . . . . . . . . 56
Organelle Lights Nuc-GFP . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Minimal Essential Medium (MEM) . . . . . . . . . . . . . . . . . . . . . . 41
Other animal sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
miRNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
mirVana miRNA Inhibitors . . . . . . . . . . . . . . . . . . . . . . . . . . 128
mirVana miRNA Mimics . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
mirVana PARIS Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
MitoSOX Red Indicator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
MitoTracker dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .91
Mobile apps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Molecular Probes Handbook . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Monoclonal Antibody Labeling Kits . . . . . . . . . . . . . . . . . . . . . 107
Mouse (C57BL/6) Mesenchymal Stem Cells . . . . . . . . . . . . . . 69
Mouse cell isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
MSC differentiation kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
MSC-Qualified FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
Multiplex immunoassays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
Multiplex magnetic assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
Mycophenolic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
P
PARIS Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .133
pBAD Expression System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
PB-MAX Karyotyping Medium . . . . . . . . . . . . . . . . . . . . . . . . 51
pcDNA vectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116
phosphoELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
pHrodo dye . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
PichiaPink Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
PichiaPink Yeast Expression System . . . . . . . . . . . . . . . . . . 118
Plant cell biology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Pluripotent stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Polar tracers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Polymyxin B sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
POROS chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Positive isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
159
Index
Index
S
Precut blotting membranes . . . . . . . . . . . . . . . . . . . . . . . . . . . 144
SAIVI Antibody Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . 107
Pre-miR Precursors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
Secondary antibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
Premo Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83, 86, 88
Secure FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Primary antibodies for flow cytometry . . . . . . . . . . . . . . . . . . 102
SeeBlue Plus2 Pre-stained Standard . . . . . . . . . . . . . . . . . . 141
Primary antibodies for western blotting . . . . . . . . . . . . . . . . . 146
SeeBlue Pre-stained Standard . . . . . . . . . . . . . . . . . . . . . . . 141
Primary Antibody Search Tool . . . . . . . . . . . . . . . . . . . . . . . . . 146
SelectFX Alexa Fluor 488 ER Labeling Kit . . . . . . . . . . . . . . 93
Primary cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
SelectFX Alexa Fluor 488 Peroxisome Labeling Kit . . . . . . . 92
Prokaryotic expression systems . . . . . . . . . . . . . . . . . . . . . . . 119
SelectFX Nuclear Labeling Kit, for fixed cells
Propidium iodide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
(DAPI, SYTOX Green, 7-AAD, TO-PRO-3 iodide) . . . . . . . . . 94
Protein A (for IP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
Selection antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Protein analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112
Serum-free media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Protein detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
Sf-900 II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .47
Protein expression . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
Sf-900 III . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Protein expression custom services . . . . . . . . . . . . . . . . . . . . 115
Sharp Protein Standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
Protein expression media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Silencer siRNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
Protein Free Hybridoma Medium-II . . . . . . . . . . . . . . . . . . . . . 42
SilverQuest Silver Staining Kit . . . . . . . . . . . . . . . . . . . . . . . 142
Protein G (for IP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
SimplyBlue SafeStain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142
Protein gel electrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
Skeletal Muscle Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . 56
Protein quantitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
Skin primary cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Protein sample fractionation . . . . . . . . . . . . . . . . . . . . . . . . . . 132
Small-scale protein labeling kits . . . . . . . . . . . . . . . . . . . . . . 107
Protein sample preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
Sodium bicarbonate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Protein stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142
Stealth Select RNAi siRNA . . . . . . . . . . . . . . . . . . . . . . . . . . 126
Protein standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
Stem cell analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67, 70
Protein tag removal with proteases . . . . . . . . . . . . . . . . . . . . 131
Stem cell detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Proteins and protein conjugates . . . . . . . . . . . . . . . . . . . . . . . . 95
Stem cell differentiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
Protocol Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Stem cell reprogramming tools . . . . . . . . . . . . . . . . . . . . . . . . 64
Pulmonary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Stem cell research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Puromycin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60, 69

StemPro Accutase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52, 63
Q
Qdot nanocrystals for multicolor flow cytometry . . . . . . . . 105
Qtracker Cell Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
QuantStudio 12K Flex Real-Time PCR System . . . . . . . . . . . 67
Qubit quantitation system . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
StemPro Differentiation Kits . . . . . . . . . . . . . . . . . . . . . . . . . . 70

StemPro EZPassage Disposable Stem Cell
Passaging Tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
StemPro hESC SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
StemPro Human Adipose-Derived Stem Cell Kit . . . . . . . . . 69
R
Rat Fetal Neural Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
ReadyProbes imaging reagents . . . . . . . . . . . . . . . . . . . . . . 101
Recombinant proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148
Recovery Cell Culture Freezing Medium . . . . . . . . . . . . . . . . 40
Resins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Rhodamine phalloidin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
RNAi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
RPMI Medium 1640 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
160
StemPro CD34+ Cell Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
StemPro LipoMAX Supplement . . . . . . . . . . . . . . . . . . . . . . 69

StemPro MSC Serum-Free Medium (SFM) . . . . . . . . . . . . . . 69
StemPro MSC SFM CTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
StemPro MSC SFM XenoFree . . . . . . . . . . . . . . . . . . . . . . . . . 68
StemPro NSC SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
StemPro Rat Alk Phos Expressing Mesenchymal
Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
StemPro-34 SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
Streptavidin, Alexa Fluor 488 conjugate . . . . . . . . . . . . . . . . 106
Index
V
Streptomycin sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Violet Ratiometric Membrane Asymmetry

Probe/Dead Cell Apoptosis Kit . . . . . . . . . . . . . . . . . . . . . . . . 84
Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
ViraPower Lentiviral T-Rex System . . . . . . . . . . . . . . . . . 116
SYPRO Ruby Protein Gel Stain . . . . . . . . . . . . . . . . . . . . . . . . 142
Virtual Cell Stain Tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
SYTO 14 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Vitronectin (VTN-N) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
SYTO 59 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Voltage sensor probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
SYTO 61 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
SYTO 82 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Index
SUMO protease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
W
Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
SYTO 9 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Western blot detection and analysis . . . . . . . . . . . . . . . . . . . . 145
SYTOX Dead Cell Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
SYTOX Nucleic Acid Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Western transfer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
T cell activation/expansion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
WesternDot 625 Western Blot Kits . . . . . . . . . . . . . . . . . . . . 145
WesternBreeze Western Blot
Immunodetection Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145

T7 expression systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
Tali Image-Based Cytometer . . . . . . . . . . . . . . . . . . . . . . . 55, 99
X
XCell II Blot Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
TAP growth media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
XCell SureLock Mini-Cell . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
TaqMan iPSC Sendai Detection Kit . . . . . . . . . . . . . . . . . . . . . 65
XCell4 SureLock Midi-Cell . . . . . . . . . . . . . . . . . . . . . . . . . . 139
TaqMan Protein Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

TC-FlAsH II tetracysteine-based protein detection . . . . . . . 88
TC-ReAsH II tetracysteine-based protein detection . . . . . . . 88
Y
Yeast expression system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118
Terrific Broth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Yeast media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Tissue Extraction Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133

TO-PRO-3 Iodide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Transfection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
trc Expression System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
T-REx Inducible Expression System . . . . . . . . . . . . . . . . . . 116
Tris-glycine gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
TrypLE Express . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Z
Zenon IgG Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
Zeocin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
ZOOM IEF Fractionator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
Zymogram gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
TrypLE Select . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
TrypLE Select CTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Trypsin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
TSA Kit #22, with HRP-streptavidin and
Alexa Fluor 488 tyramide . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
TubulinTracker Green . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
U
UltraPure 0.5 M EDTA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Umbilical Cord Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . 56
161
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Cell biology resources

2013 Life Technologies

Cell biology product and resource guide

Cell culture and cells (Chapter 2)

Cell analysis (Chapter 3)

Recovery Freezing Medium, page 40

Dynabeads cell isolation technology, page 76

Expi293 Expression System, page 42

Click-iT EdU cell proliferation assay, page 82

Classical Gibco media: DMEM, RPMI, and MEM,

DynaMag magnets, page 80

cytometry, page 102

Qdot nanocrystals for flow cytometry, page 105

pHrodo indicators, page 87

Countess Automated Cell Counter, page 53

Tali Image-Based Cytometer, page 99

Cell biology resources

Cell biology products

Protein analysis (Chapter 4)

Champion pET Expression System, page 119

Ambion Pre-miR Precursors and Anti-miR Inhibitors,

Qubit Fluorometer, page 134

Bolt and NuPAGE protein gels, page 136

SYPRO Ruby Protein Gel Stain, page 142

Novex pour-your-own gel essentials, page 136

XCell SureLock Protein Electrophoresis Cells, page 139

Cell biology research and pathways

Cell biology resources

Cell culture and cells

For Research Use Only. Not for use in diagnostic procedures.

In vitro Diagnostic Use

CTS brand research use

Class II medical device

Cell biology research and pathways

Stem cell research

Plant cell biology research

Cellular signaling pathwaysRefer to these publication-supported

Custom Novex multiplex assays for Luminex technology

Custom biology services

Website resourcesAll of these resources are available at a click, including

Cell Line Database

The Molecular Probes Handbook online

Mobile applications and widgets

Virtual Cell Staining Tool

Antibody and immunoassay selection guides

Customer and technical support

Cell biology research and pathways

Cells and culture tools for neurobiology

Rat cortex and hippocampus neuronal cells

Primary rat hippocampus neurons

Human neural stem

Media and supplements

Cell biology research and pathways

Optimized cell type

Primary rat cortex