Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb #4991

Applications Reactivity Sensitivity MW (kDa) Isotype
W IHC-P H M Endogenous 230 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse
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 (Tyr951) (15D2) Rabbit Monoclonal Antibody detects endogenous levels of VEGF receptor 2 only when phosphorylated at Tyr951. The antibody may slightly cross-react with activated VEGF receptor 1, but not with other related tyrosine phosphorylated tyrosine kinases.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr951 of human VEGF receptor 2.

Western Blotting

Western Blotting

Western blot analysis of recombinant human GST-VEGF Receptor 2 (Val789-Val1356), untreated or λ phosphatase-treated, using Phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb (upper) and VEGF Receptor 2 Antibody # 2472 (lower).

Western Blotting

Western Blotting

Phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb specifically binds to phosphorylated VEGFR2, but not other phosphorylated tyrosine kinases. Western blot analysis of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-VEGF Receptor-2 (Tyr951) (15D2) Rabbit mAb (upper) or Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (lower). CKR/PAE cells (lanes 13 and 14) express chimeric receptors containing human CSF-1 extracellular binding domain/mouse VEGF receptor-2 intracellular domain (5). CSF-1 stimulates phosphorylation of Tyr951 of intracellular VEGF receptor-2 domain (lane 13) , which was specifically detected by Phospho-VEGF Receptor-2 (Tyr951) (15D2) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HUVEC cells, untreated (left) or VEGF-treated (right), using Phospho-VEGF Receptor-2 (Tyr951) (15D2) Rabbit mAb.


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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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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

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