Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1922Y] to Cortactin
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-Cortactin antibody [EP1922Y]
See all Cortactin primary antibodies
DescriptionRabbit monoclonal [EP1922Y] to Cortactin
Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Cortactin aa 1-100 (N terminal). The exact sequence is proprietary.
- WB: HeLa and HEK-293T cell lysate. IHC-P: Human breast carcinoma. ICC/IF: MCF7 and wild-type HAP1 cells. IP: HeLa cell lysate
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
- Anti-Cortactin antibody [EP1922Y] (Alexa Fluor® 488) (ab202648)
- Anti-Cortactin antibody [EP1922Y] (Alexa Fluor® 647) (ab202650)
- Anti-Cortactin antibody [EP1922Y] (PE) (ab209758)
- Anti-Cortactin antibody [EP1922Y] (Alexa Fluor® 555) (ab215298)
- Anti-Cortactin antibody [EP1922Y] (Alexa Fluor® 594) (ab215300)
- Anti-Cortactin antibody [EP1922Y] - BSA and Azide free (ab223125)
KO cell lines
KO cell lysates
Our Abpromise guarantee covers the use of ab81208 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/50000 - 1/100000. Predicted molecular weight: 62 kDa.|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Heat mediated antigen retrieval with citrate buffer, pH 6 or Tris-EDTA, pH 9 are both suitable methods IHC-P with this antibody.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionContributes to the organization of the actin cytoskeleton and cell structure. Plays a role in the regulation of cell migration. Plays a role in the invasiveness of cancer cells, and the formation of metastases.
Sequence similaritiesContains 7 cortactin repeats.
Contains 1 SH3 domain.
DomainThe SH3 motif may mediate binding to the cytoskeleton.
modificationsTyrosine phosphorylation in transformed cells may contribute to cellular growth regulation and transformation.
Cellular localizationCytoplasm > cytoskeleton. Cell projection > lamellipodium. Cell projection > ruffle. Associated with membrane ruffles and lamellipodia.
- Information by UniProt
- Amplaxin antibody
- CTTN antibody
- EMS 1 antibody
All lanes : Anti-Cortactin antibody [EP1922Y] (ab81208) at 1/50000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CTNN knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?
Lanes 1-2: Merged signal (red and green). Green - ab81208 observed at 70 kDa. Red - loading control ab8245 observed at 37 kDa.
ab81208 Anti-Cortactin antibody [EP1922Y] was shown to specifically react with Cortactin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266819 (knockout cell lysate ab257147) was used. Wild-type and Cortactin knockout samples were subjected to SDS-PAGE. ab81208 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 50000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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.
ab81208 staining Cortactin in wild-type HAP1 cells (top panel) and Cortactin knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab81208 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
All lanes : Anti-Cortactin antibody [EP1922Y] (ab81208) at 1/2000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : Cortactin knockout HAP1 cell lysate
Lane 3 : NIH3T3 cell lysate
Lane 4 : A431 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 62 kDa
Lanes 1 - 4: Merged signal (red and green). Green - ab81208 observed at 62 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab81208 was shown to specifically react with Cortactin in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when Cortactin knockout samples were examined. Wild-type and Cortactin knockout samples were subjected to SDS-PAGE. ab81208 and ab8245 (loading control to GAPDH) were both diluted 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Purified ab81208 at 1/50 dilution (2µg) immunoprecipitating Cortactin in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab81208 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab81208 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 62 kDa
Anti-Cortactin antibody [EP1922Y] (ab81208) at 1/100000 dilution + HeLa cell lysate at 10 µg
HRP labelled Goat anti-Rabbit at 1/2000 dilution
Predicted band size: 62 kDa
Immunohistochemical analysis of paraffin-embedded human breast carcinoma staining Cortacin with ab81208 at 1/100 dilution. Heat mediated antigen retrieval with Tris-EDTA (pH 9) was perfomed.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunofluorescent staining of MCF7 cells using 1/100 ab81208
Overlay histogram showing HeLa cells stained with ab81208 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab81208, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
ab81208 has been referenced in 19 publications.
- Li Y et al. The HBx-CTTN interaction promotes cell proliferation and migration of hepatocellular carcinoma via CREB1. Cell Death Dis 10:405 (2019). PubMed: 31138777
- van de Willige D et al. Cytolinker Gas2L1 regulates axon morphology through microtubule-modulated actin stabilization. EMBO Rep 20:e47732 (2019). PubMed: 31486213
- McGuire CM et al. Isoform-specific gene disruptions reveal a role for the V-ATPase subunit a4 isoform in the invasiveness of 4T1-12B breast cancer cells. J Biol Chem 294:11248-11258 (2019). PubMed: 31167791
- Tiek DM et al. Estrogen-related receptor ß activation and isoform shifting by cdc2-like kinase inhibition restricts migration and intracranial tumor growth in glioblastoma. FASEB J 33:13476-13491 (2019). PubMed: 31570001
- Wu W et al. ß-hCG promotes epithelial ovarian cancer metastasis through ERK/MMP2 signaling pathway. Cell Cycle 18:46-59 (2019). PubMed: 30582718