Phospho-VEGF Receptor 2 (Tyr12

2477:

Western Blotting

Phospho-VEGF Receptor-2 (Tyr1212) (11A3) Rabbit mAb detects transfected levels of VEGFR-2 only when phosphorylated at tyrosine 1212. The antibody does not cross-react with other phosphorylated receptor tyrosine kinases.

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1214 of human VEGF receptor-2.

Western Blotting

Western blot analysis of recombinant human GST-VEGF Receptor 2 (Val789-Val1356), untreated or λ phosphatase-treated, using Phospho-VEGF Receptor 2 (Tyr1212) (11A3) Rabbit mAb (upper) and VEGF Receptor 2 Antibody #2472 (lower).

IP

Phospho-VEGF Receptor 2 (Tyr1212) (11A3) 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 (Tyr1212) (11A3) Rabbit mAb (upper) and Phospho-Tyrosine mAb (P-Tyr-100) #9411 (lower). CKR/PAE cells (lanes 12 and 13) express chimeric receptors containing human CSF-1 extracellular binding domain/mouse VEGF receptor-2 intracellular domain (5). CSF-1 stimulates phosphorylation of Tyr1212 of intracellular VEGF receptor-2 domain (lane 12) , which was specifically detected by Phospho-VEGF Receptor-2 (Tyr1212) (11A3) Rabbit mAb.

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.

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• 2904 VEGF Receptor 2 Control Proteins
• 8065 Human Vascular Endothelial Growth Factor-165 (hVEGF₁₆₅ )
• 3770 Phospho-VEGF Receptor 2 (Tyr1175) (D5B11) Rabbit mAb
• 2478 Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb
• 2479 VEGF Receptor 2 (55B11) Rabbit mAb
• 3817 Phospho-VEGF Receptor 2 (Tyr1059) (D5A6) Rabbit mAb
• 4991 Phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb
• 2471 Phospho-VEGF Receptor 2 (Tyr951) Antibody
• 2472 VEGF Receptor 2 Antibody
• 2474 Phospho-VEGF Receptor 2 (Tyr996) Antibody
• 2476 Phospho-VEGF Receptor 2 (Tyr951) (7H11) Mouse mAb
• 2893 VEGF Receptor 1 Antibody
• 2638 VEGF Receptor 3 (C28G5) Rabbit mAb
• 2485 VEGF Receptor 3 Antibody
• 2463 VEGF-B Antibody
• 2445 VEGF-C Antibody
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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

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