Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-FAK (Tyr925) Antibody #3284

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC H (M) (R) (C) Endogenous 125 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  C=Chicken
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Phospho-FAK (Tyr925) Antibody detects endogenous levels of FAK only when phosphorylated at tyrosine 925.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr925 of human FAK. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or anti-CD3 antibody-treated (1 µg/ml for 10 minutes), using Phospho-FAK (Tyr925) Antibody (upper) or FAK antibody (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of A549 cells, untreated (left) and lambda phosphatase-treated (right), using Phospho-FAK (Tyr925) Antibody (green). Red pseudocolor = PI/Rnase Staining Buffer® (fluorescent DNA dye).

Background

Focal adhesion kinase (FAK) is a widely expressed cytoplasmic protein tyrosine kinase involved in integrin-mediated signal transduction. It plays an important role in the control of several biological processes, including cell spreading, migration and survival (1). Activation of FAK by integrin clustering leads to autophosphorylation at Tyr397, which is a binding site for the Src family kinases PI3K and PLCγ (2-5). Recruitment of Src family kinases results in the phosphorylation of tyrosine residues 407, 576 and 577 in the catalytic domain, and tyrosine residues 871 and 925 in the carboxy-terminal region of FAK (6,7).

Phosphorylation of Tyr925 creates a binding site for the Grb2/SH2 domain and triggers a Ras-dependent activation of the MAP kinase pathway (7).

  1. Parsons, J.T. et al. (2000) Oncogene 19, 5606-5613.
  2. Schaller, M.D. et al. (1994) Mol. Cell. Biol. 14, 1680-1688.
  3. Cobb, B.S. et al. (1994) Mol. Cell. Biol. 14, 147-155.
  4. Chen, H.C. et al. (1996) J. Biol. Chem. 271, 26329-26334.
  5. Zhang, X. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 9021-9026.
  6. Calalb, M.B. et al. (1995) Mol. Cell. Biol. 15, 954-963.
  7. Schlaepfer, D.D. et al. (1994) Nature 372, 786-791.

Application References

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Companion Products

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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