Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA Antibody Pair #7824

Kit Includes Volume Cap Color
VEGFR-2 Capture Ab (100X) 0.4 ml Pink
VEGFR-2 (Y1175) Detection Ab (100X) 0.4 ml Blue
Anti-rabbit IgG, HRP-linked Antibody 0.04 ml Red

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Description

CST's PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA Kit #7335. Capture and detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The VEGFR-2 capture antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Phospho-VEGFR-2 (Tyr1175) detection antibody and anti-rabbit IgG, HRP conjugated antibody. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of Phospho-VEGFR-2 (Tyr1175) protein.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

The relationship between the protein concentration of lysate from HUVE cells, untreated or treated with VEGF, and the absorbance at 450 nM using PathScan® Phospho-VEGFR-2 (Tyr1175) Sandwich ELISA Antibody Pair #7824 is shown. HUVE cells (85% confluence) were treated with VEGF (50 ng/ml) for 5 min. at 37ºC, and then lysed.

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

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

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

Products