Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human FAK aa 700-800.
WB and ICC: HeLa
IHC: human hepatocellular carcinoma
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Non-receptor protein-tyrosine kinase implicated in signaling pathways involved in cell motility, proliferation and apoptosis. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly. Plays a potential role in oncogenic transformations resulting in increased kinase activity.
Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily. Contains 1 FERM domain. Contains 1 protein kinase domain.
The first Pro-rich domain interacts with the SH3 domain of CRK-associated substrate (BCAR1) and CASL. The carboxy-terminal region is the site of focal adhesion targeting (FAT) sequence which mediates the localization of FAK1 to focal adhesions.
Phosphorylated on 6 tyrosine residues upon activation. Microtubule-induced dephosphorylation at Tyr-397 could be catalyzed by PTPN11 and regulated by ZFYVE21. Dephosphorylated by PTPN11 upon EPHA2 activation by its ligand EFNA1.
ab40794 staining FAK in PANC-1 cells treated with CCK Octapeptide sulfated (ab120209), by ICC/IF. Increase of FAK expression correlates with increased concentration of CCK Octapeptide sulfated, as described in literature. The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120209 (CCK Octapeptide sulfated) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab40794 (1/200) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Western blot - Anti-FAK antibody [EP695Y] (ab40794)
Anti-FAK antibody [EP695Y] (ab40794) at 1/1000 dilution + Hela cell lysate
Predicted band size : 119 kDa Observed band size : 119 kDa
Immunofluorescent staining of HeLa cells using ab40794.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP695Y] (ab40794)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom
The image shows FAK antibody (ab40794) in human spleen tissue. Clear cytoplasmic positivity in a subset of germinal centre cells.
The there is intense positivity of the serum in the blood vessels. Endogenous peroxidases was blocked using 2% H2O2, for 15 minutes.
Overlay histogram showing HeLa cells stained with ab40794 (red line). The cells were fixed with 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 (ab40794, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.
Western blot - Anti-FAK antibody [EP695Y] (ab40794)This image is courtesy of an Abreview submitted by Magali Boissiere (6970246)
All lanes : Anti-FAK antibody [EP695Y] (ab40794)
Lane 1 : Milk PBS Tween Lane 2 : Milk PBS Tween
Blocking peptides at 5 % per lane.
Secondary HRP conjugated goat anti-rabbit poly clonal at 1/5000 dilution Developed using the ECL technique
This image is courtesy of an Abreview submitted by Magali Boissiere (6970246)
Western blot analysis of RAW264.7 cells lysate (40μg/lane) labelling FAK with ab40794 at 1/5000 in 5% Milk PBS Tween for 16 hours at 4ºC. A HRP conjugated goat anti-rabbit poly clonal (1/5000) was used as the secondary antibody.
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