Product datasheet

Anti-Cytokeratin 8 antibody [EP1628Y] ab53280

Recombinant

3 Abreviews 24 References 12 Images

Overview

Product name Anti-Cytokeratin 8 antibody [EP1628Y]

Description Rabbit monoclonal [EP1628Y] to Cytokeratin 8
Tested applications Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IF
Species reactivity Reacts with: Mouse, Human
Immunogen Synthetic peptide within Human Cytokeratin 8 aa 300-400 (C terminal). The exact sequence is
proprietary.
Database link: P05787

Positive control A431 cell lysate, HeLa cells or human breast adenocarcinoma tissue.
General notes This product is a recombinant rabbit monoclonal antibody.
A trial size is available to purchase for this antibody.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these
species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit
monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

Form Liquid
Storage instructions Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Dissociation constant (KD) KD = 4.60 x 10 -10 M

Learn more about KD

Storage buffer PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%

Purity Tissue culture supernatant
Clonality Monoclonal
Clone number EP1628Y
1
Isotype IgG

Applications

Our Abpromise guarantee covers the use of ab53280 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes

WB 1/25000 - 1/50000. Detects a band of approximately 52 kDa (predicted molecular weight: 54 kDa).

IP 1/70.

Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration.

ICC/IF 1/100 - 1/500.

Target

Function Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of
striated muscle.
Tissue specificity Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma
membrane in structures that contain dystrophin and spectrin. Expressed in gingival mucosa and
hard palate of the oral cavity.
Involvement in disease Cirrhosis
Sequence similarities Belongs to the intermediate filament family.
Post-translational Phosphorylation on serine residues is enhanced during EGF stimulation and mitosis. Ser-74
modifications phosphorylation plays an important role in keratin filament reorganization.
O-glycosylated. O-GlcNAcylation at multiple sites increases solubility, and decreases stability by
inducing proteasomal degradation.
O-glycosylated (O-GlcNAcylated), in a cell cycle-dependent manner.
Cellular localization Cytoplasm. Nucleus, nucleoplasm. Nucleus matrix.

Images

2
 Image: Courtesy of Dr. Shaohua Li, UMDNJ-
Robert Wood Johnson Medical School

Sample: mouse embryonic stem cell-

differentiated embryoid bodies (EBs)

Preparation:

Fix in 3%PFA in PBS for 30 min at

RTIncubate in 7.5% sucrose-PBS for 3h at
RTIncubate in 15% sucrose-PBS at 4 degree
Celsius overnightEmbed the EBs in tissue-
Immunohistochemistry (Formalin/PFA-fixed paraffin-
Tek OCT compoundCut frozen sections to 4-
embedded sections) - Anti-Cytokeratin 8 antibody
20 µm thickness
[EP1628Y] (ab53280)
This image is courtesy of Dr. Shaohua Li
Primary antibody 1: Rabbit anti cytokeratin 8
(ab53280), 1:100

Primary antibody 2: Rat anti-perlecan, 1:100

Secondary antibody 1: Goat polyclonal
Secondary Antibody to Rabbit IgG - H&L
(Alexa Fluor® 488) pre-adsorbed (ab150081),
1:200

Secondary antibody 2: Goat polyclonal

Secondary Antibody to Rat IgG - H&L (Cy5®)
pre-adsorbed (ab150081), 1:200
Nuclei were counterstained with DAPI

3
 Immunocytochemistry/Immunofluorescence
analysis of HeLa (human cervix
adenocarcinoma) labelling Cytokeratin 8 with
purified ab53280 at 1/500. Cells were fixed
with 4% Paraformaldehyde and permeabilised
by 0.1% tritonX-100. An Alexa Fluor® 488-
conjugated goat anti-rabbit IgG (1/1000) was
used as the secondary antibody (Ab150077).
Nuclei counterstained with DAPI (blue).

Immunocytochemistry/ Immunofluorescence - Anti-

Cytokeratin 8 antibody [EP1628Y] (ab53280)

Ab53280 (1:250) staining human Cyotkeratin

8 in human breast adenocarcinoma tissue by
immunohistochemistry using paraffin
embedded tissue.

Immunohistochemistry (Formalin/PFA-fixed paraffin-

embedded sections) - Anti-Cytokeratin 8 antibody
[EP1628Y] (ab53280)

Anti-Cytokeratin 8 antibody [EP1628Y]

(ab53280) at 1/50000 dilution + A431 cell
lysate at 10 µg

Secondary
Goat anti-Rabbit HRP labeled at 1/2000
dilution

Predicted band size : 54 kDa

Observed band size : 52 kDa

Western blot - Anti-Cytokeratin 8 antibody

[EP1628Y] (ab53280)

4

Immunohistochemistry (Formalin/PFA-fixed paraffin-
embedded sections) - Anti-Cytokeratin 8 antibody
[EP1628Y] (ab53280)
This image is courtesy of an anonymous Abreview
Immunocytochemistry/ Immunofluorescence - Anti-
Cytokeratin 8 antibody [EP1628Y] (ab53280)
Immunohistochemistry (Formalin/PFA-fixed paraffin-
embedded sections) - Anti-Cytokeratin 8 antibody
[EP1628Y] (ab53280)
ab53280 staining Cytokeratin 8 in Mouse liver
tissue sections by Immunohistochemistry (IHC-
P - paraformaldehyde-fixed, paraffin-
embedded sections). Tissue was fixed with
formalin and blocked with 10% serum for 20
minutes at 23°C; antigen retrieval was by heat
mediation in a citrate buffer. Samples were
incubated with primary antibody (1/75 in TBS
+ 1% BSA) for 1 hour at 23°C. A Biotin-
conjugated Goat anti-rabbit IgG polyclonal
(1/500) was used as the secondary antibody.
Immunofluorescent staining of HeLa cells
using ab53280 (1:100).
Fluorescent immunohistochemical analysis of
paraffin-embedded human ovarian carcinoma
tissue using ab53280. Green-CK8 red-PI.
5
 Fluorescent immunohistochemical analysis of
paraffin-embedded human breast carcinoma
tissue using ab53280. Green-CK8 red-PI

Immunohistochemistry (Formalin/PFA-fixed paraffin-

embedded sections) - Anti-Cytokeratin 8 antibody
[EP1628Y] (ab53280)

Fluorescent immunohistochemical analysis of

paraffin-embedded human colonic
adenocarcinoma tissue using ab53280.
Green-CK8 red-PI.

Immunohistochemistry (Formalin/PFA-fixed paraffin-

embedded sections) - Anti-Cytokeratin 8 antibody
[EP1628Y] (ab53280)

Fluorescent immunohistochemical analysis of

paraffin-embedded human endometrial
carcinoma tissue using ab53280. Green-CK8
red-PI.

Immunohistochemistry (Formalin/PFA-fixed paraffin-

embedded sections) - Anti-Cytokeratin 8 antibody
[EP1628Y] (ab53280)

6

Flow Cytometry - Anti-Cytokeratin 8 antibody
[EP1628Y] (ab53280)
Overlay histogram showing HeLa cells stained
with ab53280 (red line). The cells were fixed
with 2% PFA (room temperature, 30 min) and
then permeabilized with 1% FACS
permeabilizing solution for 30 min. The cells
were then incubated in 3% FBS in 1X
PBS followed by the antibody (ab53280, 1/20
dilution) for 1 hour at room temperature. The
cells were then incubated for 30 min at room
temperature with the secondary antibody. An
isotype control antibody (black line) was used
and an unlabelled sample (blue line) was also
used as a control.

Equilibrium disassociation constant (KD)

Learn more about KD

Click here to learn more about KD

Other - Anti-Cytokeratin 8 antibody [EP1628Y]

(ab53280)

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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