SpecificityPhosphorylation site-specific antibody selective for the phosphorylated form of the Focal Adhesion Kinase enzyme containing a phosphate on tyrosine 407. The antibody has been shown to recognize Focal Adhesion Kinase (approximately 125 kDa) in chick embryo fibroblast cells plated on fibronectin.
Synthetic peptide derived from the region of Focal Adhesion Kinase that contains tyrosine 407. The sequence is conserved in human, mouse, rat, chicken and frog.
Focal Adhesion Kinase is a non-receptor protein tyrosine kinase discovered as a substrate for Src and as a key element of integrin signalling. Focal Adhesion Kinase plays a central role in cell spreading, differentiation, migration, cell death and acceleration of the G1 to S phase transition of the cell cycle. The activity of the phosphorylation site pTyr407 is currently unknown.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Purification notesPurified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using (i) a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Focal Adhesion Kinase enzyme and (ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphotyrosine, irrespective of the sequence. The final product is generated by affinity chromatography using a Focal Adhesion Kinase-derived peptide that is phosphorylated at tyrosine 407.
Primary antibody notesFocal Adhesion Kinase is a non-receptor protein tyrosine kinase discovered as a substrate for Src and as a key element of integrin signalling. Focal Adhesion Kinase plays a central role in cell spreading, differentiation, migration, cell death and acceleration of the G1 to S phase transition of the cell cycle. The activity of the phosphorylation site pTyr407 is currently unknown.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Predicted molecular weight: 125 kDa.
FunctionNon-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.
Tissue specificityExpressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily. Contains 1 FERM domain. Contains 1 protein kinase domain.
DomainThe 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.
Post-translational modificationsPhosphorylated 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.
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Anti-FAK (phospho Y407) antibody images
Western blot - FAK (phospho Y407) antibody (ab4814)
Predicted band size : 125 kDa
Cell extracts prepared from chick embryo fibroblasts expressing FAK and plated on fibronectin were resolved by SDS-PAGE on a 4-20% Tris-glycine gel. The proteins were then transferred to nitrocellulose. Membranes were incubated with 0.3 µg/mL ab4814, following prior incubation in the absence (none) or presence of the peptide immunogen, the non-phosphopeptide corresponding to the FAK phosphopeptide, or a generic phosphotyrosine peptide. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar detection method. The data show that only the phosphopeptide corresponding to this site blocks the antibody signal, therefore demonstrating the specificity of ab4814 for this phosphorylated residue.