Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-VEGF Receptor 2 (Tyr996) Antibody #2474

Applications Reactivity Sensitivity MW (kDa) Source
W H M Transfected Only 230 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Phospho-VEGF Receptor 2 (Tyr996) Antibody detects transfected VEGFR 2 only when phosphorylated at tyrosine 996. The antibody cross-reacts with tyrosine-phosphorylated PDGF receptor.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr996 of human VEGF receptor 2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from CKR/PAE cells expressing chimeric receptors containing human CSF-1 extracellular binding domain/mouse VEGF receptor 2 intracellular domains (Rahimi, N. et al. (2000) J. Biol. Chem. 275, 16986-16992), using Phospho-VEGF Receptor 2 (Tyr996) Antibody (upper) or VEGF receptor 2 antibody (lower).

Background

Vascular endothelial growth factor receptor 2 (VEGFR2, KDR, Flk-1) is a major receptor for VEGF-induced signaling in endothelial cells. Upon ligand binding, VEGFR2 undergoes autophosphorylation and becomes activated (1). Major autophosphorylation sites of VEGFR2 are located in the kinase insert domain (Tyr951/996) and in the tyrosine kinase catalytic domain (Tyr1054/1059) (2). Activation of the receptor leads to rapid recruitment of adaptor proteins, including Shc, GRB2, PI-3 kinase, Nck and the protein tyrosine phosphatases SHP-1 and SHP-2 (3). The phosphorylation of Tyr1212 provides a docking site for Grb2 binding and phospho-Tyr1175 binds with the p85 subunit of PI-3 kinase and PLCγ, as well as Shb (1,4,5). Signaling from VEGFR2 is necessary for the execution of VEGF-stimulated proliferation, chemotaxis and sprouting, as well as survival of cultured endothelial cells in vitro and angiogenesis in vivo (6-8).

  1. Meyer, M. et al. (1999) EMBO J. 18, 363-374.
  2. Dougher-Vermazen, M. et al. (1994) Biochem. Biophys. Res. Commun. 205, 728-738.
  3. Kroll, J. and Waltenberger, J. (1997) J. Biol. Chem. 272, 32521-32527.
  4. Takahashi, T. et al. (2001) EMBO J. 20, 2768-2778.
  5. Holmqvist, K. et al. (2004) J. Biol. Chem. 279, 22267-22275.
  6. Karkkainen, M.J. and Petrova, T. (2000) Oncogene 19, 5598-5605.
  7. Rahimi, N. et al. (2000) J. Biol. Chem. 275, 16986-16992.
  8. Claesson-Welsh, L. (2003) Biochem. Soc. Trans. 31, 20-24.

Application References

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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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