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Technical Data Sheet

FM1211

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FAK (Tyr-397), phospho-specific
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms, Rb
125 kDa

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Western blot analysis of HUVECs untreated (lanes 1 & 3) or treated with alkaline phosphatase (lanes 2 & 4). Blots were probed with mouse monoclonal anti-FAK (lanes 1 & 2) and anti-FAK (Tyr-397) (lanes 3 & 4).

Application
Dilution


ELISA
1:2000


WB
1:500



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
Creekmore, A.L. et al. (2013) Biomolecules 3(3): 386. WB: Mouse cells
Teravainen, T.P. et al. (2013) PLoS ONE 8(8):e71485. WB: MDCK cells
Lin, S. & Mequanint, K. (2012) Biomaterials 33(29):7047. WB: HCASMC
Akama, K. et al. (2011) BBA Prot & Proteo. 1814(2):265. WB: monkey neurons
Kong, T. et al. (2009) Mol Biol Cell. 20:4596. WB: canine MDCK cells
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Background
Focal adhesion kinase (FAK) is a widely expressed cytoplasmic protein tyrosine kinase involved in signal transduction pathways important for cell spreading, migration and survival. Activation of FAK by integrin clustering leads to autophosphorylation at Tyr-397, which is a binding site for Src family kinases, PI3-Kinase, and PLCγ. The recruitment of Src family kinases results in the phosphorylation of tyrosine 407, 576, and 577 in the catalytic domain, and tyrosine 871 and 925 in the carboxy-terminal region of FAK. Thus, the phosphorylation of Tyr-397 is a critical step in the activation of FAK.


Background References
Cobb, B.S. et al. (1994) Mol. Cell. Biol. 14:147.
Schaller, M.D. et al. (1994) Mol. Cell. Biol. 14:1680.
Schlaepfer, D.D. et al. (1994) Nature 372:786-791.
Parsons, J.T. et al. (2000) Oncogene 19:5606.
Immunogen
Clone (M121) was generated from a synthetic peptide (coupled to KLH) corresponding to amino acid residues around tyrosine 397 of human FAK. This peptide sequence has high homology to the conserved tyrosine site in rat and mouse FAK.
Buffer and Storage
Mouse monoclonal antibody purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
The antibody detects a 125 kDa* protein on SDS-PAGE immunoblots of untreated HUVEC cells. This phosphorylated band is greatly reduced after treatment with alkaline phosphatase.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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