Simple Western

Technology Overview
Yasmine Aurellia
Product Specialist
 Meet Bio-Techne!
Proteins are Their Core

• Quantikine ELISAs • Primary Antibodies • Protein Characterization in a

• DuoSet ELISAs
• Luminex
• Primary Antibodies
• Secondary Antibodies
• Proteins
• Secondary Antibodies Capillary
• IHC Tissue Slides Ella: Simple Plex
• IHC Tissue Microarrays Jess, Abby: Simple Western
Maurice: Simple icIEF + CE-
SDS
MFI: Simple Particle Analysis
Do Your Westerns Look Like This?

• Manual, multi-step process.

Highly prone to errors
• Inconsistent. Same assay,
different results
• Time consuming, 24 hours+
Traditional Western
More Steps, More Errors

• Up to 18 steps.
Multiple reagent
additions and wash
steps
• Each step introduces
an opportunity for
error
• Error equals
inconsistency
 Meet Jess
Automated Western Blot Systems

✔ No gels, no transfer devices, no

blots, no film, no manual steps
are involved
✔ Fully analyzed results in 3 hours
✔ Save cost, time, and resources
 Transforming Complexity into Simplicity:
Assay Set-Up is Easy

Insert Capillary
Cartridge

Prepare Samples and

Reagents

Automatically
Press Start
Analyzed Results
Jess Instruments Operations
Meet the
Simple Western Family!
The Only Fully Automated Western Device on the Market
 Peggy Sue
Uncover Their Unique Sally Sue NanoPro 1000

Features
Jess Abby

3 hrs. 14 hrs. (size)

14 hrs. 11 hrs.
5 hrs. (RePlex) 11 hrs. (charge)

12-24 samples 12-96 samples

Size Size Size or Charge Charge

Chemi. &
Chemi. Chemi.
Fluor.

RePlex N/A

Blot Imaging N/A

Simple Western
Outperforms Traditional
Western in Every Aspect

Jess, Abby
Assay Parameter Traditional Western
(Simple Western)
Hands-on Time 30 Minutes 3 Hours
Time to Result 3 Hours 3 Days
Sample Volume 3 µL 10-50 µL
Dynamic Range 3-4 Logs ~2.5 Logs
LOD 1 pg/µL 3.4 pg/µL
Sensitivity Low pg Low ng
Quantitative Quantitative Semi-Quantitative
Time Savings Summary
Hands-on Time Time to Your
Throughput
Result Freedom

30 3 Up to 75
Jess, Abby Increased
Minutes Hours Samples
Per Day

Sally Sue, Peggy 45 11–14 96 Samples Increased

Sue, NanoPro1000 Minutes Hours Per Day

3 24-72
Traditional Western 20 Confined
Hours Hours
Simple Western
Gel-Free, Transfer-Free,
Blot-Free, Hands-Free
Unlike a traditional western,
there is no:
• Gel or apparatus
preparation
• Transfer to a membrane
• Manual incubation or wash
steps
• Developing/wasting film
• Subjective data analysis

All Processes Take

Place in a Capillary
Separation Kits
• 2-40kDa separation range
• 12-230kDa separation range
• 66-440kDa separation range

Its All in the Kit

Wash Buffer
Standard

Capillary Cartridge
Pre-filled Plate

Sample Buffer
Detection Kits
• Chemiluminescence
• Fluorescence

Its All in the Kit

Luminol-S
Streptavidin-HRP
Peroxide
Antibody Diluent
Secondary Antibody
Capillary Cartridge

• Available 13 and 25 capillaries

• Vertical separation

• Capillary cartridge captures all

waste
Pre-filled Plate

• Separation components pre-filled

• Contains all liquids such as
separation matrix, stacking
matrix, matrix removal buffer,
upper running buffer, lower
running buffer
Blot Cartridge

Perfect all-in-one solution for traditional

Western Blot imaging
• Chemiluminescence based
• Automatically detected in Jess
• Image, exposure adjustment, contrast adjustment
• Export out of Compass for analysis
How it Works:
Only 3 Simple
Steps!

3. Run
• Insert Plate
2. Dispense • Insert Capillary
• Pipette Ladder Cartridge
• Pipette Sample • Start Run
• Pipette Ab Diluent • Magic Happens in the
• Pipette 1st Ab Capillary
1. Prepare Reagents
• Pipette 2nd Ab
• Pipette
• Mix • Pipette Detection Quick + Easy
• Heat • Wash Buffer = Results
In 3 Hours Later..
Get Multiple Views of Results

Capillary Image Electropherogram Lane View

View View
 21 pg AKT/µL sample

Endogenous
AKT
Truly Quantitative AKT-GST
Standard
Curve
Share Your Data with Confidence
Standardized output and system controls allow quantitative comparison:
• Person to Person
• Lab to Lab
• Site to Site Software Integrated
Functionality for Compliance
with 21 CFR Part 11
Two Colour Fluorescence

Two Color Fluorescence

Lane View with Overlay Graph View with Intensity

Emission Wavelength Detection Kits

NIR = ~700 nm Anti-Rabbit, Anti-Mouse NIR
IR = ~800 nm Anti-Rabbit, Anti-Mouse IR
Protein Normalization

200000
180000
160000
140000

Peak Area Count

120000
100000
80000
60000
40000
20000
0
0.4mg/mL 0.6mg/ml 0.8mg/ml

Ave. Area Ave. Corr. Area

 Simple Western Makes Life Simple

• 30-Minute Workflow
Pre-filled plate
Disposable capillary cartridge
• Results in Under 3 Hours
Run up to 75 samples/day
• More Sensitive than Traditional Western
Use as little as 0.3-3.6 μg/μL protein
Requires just 3 uL/well
Your Sample is Precious
• Are you working too hard to obtain enough cells or tissue?
• Are you pooling samples together before running your Western Blot?
• Can you afford to use 10-50 µL of your sample for just one gel lane?

Traditional
Western Blot Simple
Western
10-50
µL/lane 3 µL/lane

Imagine what you could do differently

if you only needed 3 µL
 Case Study
Professor Dr. Rieger works with mouse hematopoietic stem cells
Simple Westerns assays enable him to get more data with his precious sample

Start with a mouse ($20-$500)

• Isolate bone marrow cells from thigh Professor Dr. Rieger
LOEWE Centre for Cell & Gene Therapy
• Isolate stem cells
Goethe University Hospital Frankfurt, Germany
• 1 stem cell per 7,000 bone marrow cells
• 1,000 stem cells per mouse

Traditional Western Simple Western

🗷 Requires 50 µL or 50,000 cells ✔ Requires 3 µL or 3,000 cells
🗷 Approximately 50 mice ✔ Approximately 3 mice
🗷 Estimated cost of $1000-$25,000 ✔ Estimated cost of $60-$1500
per lane
Low Volume, Big Results
Be a Sample Conservationist

Jess, Abby
Platform Traditional Western
(Simple Western)

Sample Volume 3 µL 10–50 µL

Quantitation CV <10% >35%

LOD 1 pg/μL 5 pg/μL

Simple Western Applications
Detecting Proteins in Urine

Protein 1

Protein 2

Get
numbers
Important Protein in Plant Breeding Measured Using
Simple Western Size

1
2
3

SYN1
Detection of a Component of the Photosynthetic
Photosystem II Complex in Leaf Extract
Detecting Tagged Proteins Using
Simple Western Size

His-tag FLAG-tag
PI3K Pathway Profiling Fine Needle Aspirates (FNA)
and Tissue Samples

Detecting 7 Distinct Targets in the PI3K Signaling Pathway

Similar Sensitivity Compared to Coomassie Staining
Coomassie Total
Protein 2-fold serial dilutions starting with sample 1
(*): 230kDa Fluorescent Standard
*
250-
A
150-
A A: Alpha-2-macroglobulin
B
100-
B B: Beta-galactosidase
75-
C
C C: Bovine Serum Albumin
50-

D
37- D D: Ovalbumin

E E E: Carbonic Anhydrase
25-

20-
F F F: RNaseA
15-
They Have Already Embraced the Simplicity and
Effectiveness of Simple Western
Database of Documented
Antibodies Run on Simple
Western Platform
Simple Western Antibody
Database | Bio-Techne
Simple Western certified antibodies from
R&D Syst and Novus Biologicals

Certification Means:
• Suggested antibody concentration
• Suggested lysate concentration
• Data from scientifically relevant lysate
• Suggested optimal lysate concentration
• 100% guarantee

Innovator’s Reward:
• Submit your Simple Western data on an antibody that has not
been tested yet, and receive a free Simple Western assay
module
 Interactive Training Portal
The Bio-Techne Academy is a free, comprehensive online learning platform which offers support
and training on a variety of topics and for any skill level.

✔ Free resources for all

✔ Accessible anytime, anywhere
✔ Expert instructors at your fingertips
Why Simple Western Is Your Ultimate Solution
Effortless, Efficient, Extraordinary
• Fully automated, just load
Fast Time to samples, and press start
Results • Fully analysed results in 3
hours

• Save samples, reagents,

Reduce Costs time, and labor with RePlex
kit
 High • Industry-leading sensitivity
Sensitivity in chemiluminescence and
Multiplex fluorescence

Low Sample • Use as little as 3 µL per well

Volume to get pg-sensitivity

Reproducible • Built in analysis software

and • Absolute and relative
Quantitative protein quantitation
Thank You
Questions?