Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-VEGF Receptor 2 (Tyr1059) (D5A6) Rabbit mAb #3817

Applications Reactivity Sensitivity MW (kDa) Isotype
W H M (R) Endogenous 230 Rabbit IgG

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-VEGF Receptor 2 (Tyr1059) (D5A6) Rabbit mAb only detects endogenous levels of VEGFR2 proteins when phosphorylated at Tyr1059. Since VEGF receptors 1, 2 and 3 share identical sequences within the epitope region, this antibody can also detect VEGF receptors 1 and 3 when phosphorylated at corresponding tyrosine residues.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1059 of human VEGF receptor 2.

Western Blotting

Western Blotting

Western blot analysis of PAE/CKR cells, untreated or stimulated with CSF-1, using Phospho-VEGF Receptor 2 (Tyr1059) (D5A6) Rabbit mAb (upper) and VEGF Receptor 2 (55B11) Rabbit mAb #2479 (lower). PAE/CKR cells express a chimeric receptor made up of human CSF-1 receptor extracellular domain and mouse VEGF receptor 2 transmembrane and intracellular domains.

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, PI3 kinase, NCK, and the protein tyrosine phosphatases SHP-1 and SHP-2 (3). Phosphorylation at Tyr1212 provides a docking site for GRB2 binding and phospho-Tyr1175 binds the p85 subunit of PI3 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, 363-74.
  2. Dougher-Vermazen, M. et al. (1994) Biochem Biophys Res Commun 205, 728-38.
  3. Kroll, J. and Waltenberger, J. (1997) J Biol Chem 272, 32521-7.
  4. Takahashi, T. et al. (2001) EMBO J 20, 2768-78.
  5. Holmqvist, K. et al. (2004) J Biol Chem 279, 22267-75.
  6. Karkkainen, M.J. and Petrova, T.V. (2000) Oncogene 19, 5598-605.
  7. Rahimi, N. et al. (2000) J Biol Chem 275, 16986-92.
  8. Claesson-Welsh, L. (2003) Biochem Soc Trans 31, 20-4.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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