Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb #2478

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

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
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 (Tyr1175) (19A10) Rabbit mAb detects endogenous levels of VEGFR-2 proteins only when phosphorylated at tyrosine 1175. This antibody may cross-react with VEGFR1.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1175 of human VEGF receptor-2.

Western Blotting

Western Blotting

Phospho-VEGF Receptor 2 (Tyr1175) (19A10) 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 (Tyr1175) (19A10) 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 (7). CSF-1 stimulates phosphorylation of Tyr1175 of intracellular VEGF receptor-2 domain (lane 12), which was specifically detected by Phospho-VEGF Receptor-2 (Tyr1175) (19A10) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HUVE cells, untreated or stimulated with VEGF (50 ng/ml for 2 minutes), using Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb (upper) and VEGF Receptor 2 (55B11) Rabbit mAb #2479 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HUVE cells, untreated (left) or VEGF treated (right).


IF-IC

IF-IC

Confocal immunofluorescent images of HUVE cells untreated (left) or stimulated with Vascular Endothelial Growth Factor (VEGF) #9943 (right) and labeled with Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb (top, green) and VEGF Receptor 2 (55B11) Rabbit mAb #2479 (bottom, green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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, 363-74.
  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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Products