Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker is a rabbit recombinant monoclonal antibody that is used to detect Cytokeratin 8 in Flow cytometry (Intra), ICC/IF, IHC-Fr, IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.
- Specificity confirmed with KRT8 knockout cell line validation
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 130 publications
- Trusted since 2007
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IP | WB | IHC-Fr | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|---|
Human | Tested | Tested | Expected | Tested | Tested | Tested |
Mouse | Expected | Tested | Tested | Expected | Tested | Tested |
Rat | Expected | Expected | Predicted | Expected | Predicted | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/20 | Notes For unpurified use at 1:70. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species Rat | Dilution info 1/20 | Notes For unpurified use at 1:70. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/10000 | Notes For unpurified use at 1/25,000 - 1/50,000. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/10000 | Notes For unpurified use at 1/25,000 - 1/50,000. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/10000 | Notes For unpurified use at 1/25,000 - 1/50,000. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species Rat | Dilution info 1/100 - 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/20 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species Mouse | Dilution info 1/20 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/250 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/250 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/250 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle.
CYK8, KRT8, Cytokeratin-8, Keratin-8, Type-II keratin Kb8, CK-8, K8
Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker is a rabbit recombinant monoclonal antibody that is used to detect Cytokeratin 8 in Flow cytometry (Intra), ICC/IF, IHC-Fr, IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.
- Specificity confirmed with KRT8 knockout cell line validation
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 130 publications
- Trusted since 2007
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-Fr, IHC-P, IP, WB in human, mouse, rat samples.
Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) has been cited over 135 times in peer reviewed journals and is trusted by the scientific community.
Abcams high quality manufacturing and validation processes ensure Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) has been confirmed by testing in knockout samples.
Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) has 9 independent reviews from customers.
Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) specifically detects Cytokeratin 8 (UniProt ID: P05787; Molecular weight: 54kDa) and is sold in 100 uL and 1 mL selling sizes.
Conjugation-ready, carrier free format available for antibody clone EP1628Y - Anti-Cytokeratin 8 antibody [EP1628Y] - BSA and Azide free ab217173.
Antibody clone EP1628Y is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488, Alexa Fluor® 647, HRP, PE, Alexa Fluor® 45 (Alexa Fluor® 488 Anti-Cytokeratin 8 antibody [EP1628Y] ab192467, Alexa Fluor® 647 Anti-Cytokeratin 8 antibody [EP1628Y] ab192468, ab19394, Mouse lung normal tissue lysate - total protein ab29297, ab21139).
Cytokeratin 8 (CK8) is a type II cytoskeletal keratin protein that is used as a diagnostic marker to differentiate various carcinomas. CK8 is involved in cell signaling, attachment, and stress adaptation, which are crucial for cancer cell survival and progression.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Species reactivity
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling Cytokeratin 8 with Purified ab53280 at 1:250 dilution. Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
ab53280 (purified) at 1:20 dilution (0.2μg) immunoprecipitating Cytokeratin 8 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10μg
Lane 2 (+): ab53280 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab53280 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Predicted band size: 53 kDa
Cytokeratin 8 Western blot staining using rabbit Anti-Cytokeratin 8 antibody
ab53280 was shown to react with Cytokeratin 8 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human KRT8 (Cytokeratin 8) knockout HeLa cell line ab255400 (knockout cell lysate Human KRT8 (Cytokeratin 8) knockout HeLa cell lysate ab263785) was used. Wild-type and Cytokeratin 8 knockout samples were subjected to SDS-PAGE. ab53280 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 10000 (For unpurified use at 1/25,000 - 1/50,000) dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) at 1/10000 dilution
Lane 1: A431 cell lysate at 20 µg
Lane 2: MCF7 cell lysate at 20 µg
Lane 3: Wild-type HeLa cell lysate at 20 µg
Lane 4: Western blot - Human KRT8 (Cytokeratin 8) knockout HeLa cell lysate (Human KRT8 (Cytokeratin 8) knockout HeLa cell lysate ab263785)
Lane 4: Western blot - Human KRT8 (Cytokeratin 8) knockout HeLa cell line (Human KRT8 (Cytokeratin 8) knockout HeLa cell line ab255400)
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 53 kDa
Observed band size: 37 kDa, 52 kDa
Immunofluorescent analysis of luminal epithelial marker keratin 8 (green) and myoepithelial marker keratin14 (red) revealed a similar staining pattern in Panx1-/- mice compared to control mice during lactation. Paraffin-embedded tissue samples.
Hoescht (blue) denotes nuclei. N = 6. Scale bars = 50 um.
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 8 with purified ab53280 at 1/20 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Alexa Fluorr®488 (Alexa Fluor® 488 Anti-Cytokeratin 8 antibody [EP1628Y] ab192467) and Alexa Fluorr®647 (Alexa Fluor® 647 Anti-Cytokeratin 8 antibody [EP1628Y] ab192468) conjugated versions are available for this clone.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) at 1/10000 dilution
Lane 1: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 2: Human breast cancer lysates at 20 µg
Lane 3: HaCaT (Human skin keratinocyte) whole cell lysates at 20 µg
Lane 4: Mouse colon lysates at 20 µg
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 53 kDa
Immunocytochemistry/ Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling Cytokeratin 9 with Purified ab53280 at 1:500 dilution. Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Alexa Fluor® 488 (Alexa Fluor® 488 Anti-Cytokeratin 8 antibody [EP1628Y] ab192467) and Alexa Fluor® 647 (Alexa Fluor® 647 Anti-Cytokeratin 8 antibody [EP1628Y] ab192468) conjugated versions are available for this clone.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Cytokeratin 8 with Purified ab53280 at 1:250 dilution. Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
Unpurified ab53280 (1:250) staining human Cyotkeratin 8 in human breast adenocarcinoma tissue by immunohistochemistry using paraffin embedded tissue.
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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Nuclei counterstained with DAPI (blue).
All lanes: Western blot - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) at 1/50000 dilution
All lanes: A431 cell lysate at 10 µg
All lanes: Goat anti-Rabbit HRP labeled at 1/2000 dilution
Predicted band size: 53 kDa
Observed band size: 52 kDa
Overlay histogram showingHeLa cells stained with unpurifiedab53280 (red line). The cells were fixed with 2%PFA (room temperature, 30 min) and then permeabilized with 1%FACS permeabilizing solutionfor 30 min. The cells were then incubated in3% FBS in 1X PBSfollowed by the antibody (ab53280, 1/20 dilution) for1 hourat 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.
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 RT Incubate in 7.5% sucrose-PBS for 3h at RT Incubate in 15% sucrose-PBS at 4 degree Celsius overnight Embed the EBs in tissue-Tek OCT compound Cut frozen sections to 4-20 μm thickness
Primary antibody 1: Rabbit anti cytokeratin 8 (unpurified 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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081), 1:200
Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081), 1:200
Nuclei were counterstained with DAPI
Unpurified 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.
Fluorescent immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue using unpurified ab53280. Green-CK8 red-PI.
Fluorescent immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue using unpurified ab53280. Green-CK8 red-PI.
Fluorescent immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue using unpurified ab53280. Green-CK8 red-PI.
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab53280. Green-CK8 red-PI
Flow cytometry overlay histogram showing left NIH3T3 positive cells and right negative Raw264.7 stained with ab53280 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab53280) (1x 106 in 100μl at 0.2μg/ml (1/11500)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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