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 Dm | Endogenous | 230 | Rabbit IgG |
Applications Key:
W=Western Blotting
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
Dm=D. melanogaster
Species cross-reactivity is determined by Western blot.
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 antibodies are produced by immunizing animals with a synthetic phospho-peptide (KLH coupled) corresponding to residues surrounding Tyr1175 of human VEGF receptor-2.
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.
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 (5). 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.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded HUVE cells, untreated (left) or VEGF treated (right).
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™ (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).
- Meyer, M. et al. (1999) EMBO J. 18, 363-374.
- Dougher-Vermazen, M. et al. (1994) Biochem. Biophys. Res. Commun. 205, 728-738.
- Kroll, J. and Waltenberger, J. (1997) J. Biol. Chem. 272, 32521-32527.
- Takahashi, T. et al. (2001) EMBO J. 20, 2768-2778.
- Holmqvist, K. et al. (2004) J. Biol. Chem. 279, 22267-22275.
- Karkkainen, M.J. and Petrova, T. (2000) Oncogene 19, 5598-5605.
- Rahimi, N. et al. (2000) J. Biol. Chem. 275, 16986-16992.
- Claesson-Welsh, L. (2003) Biochem. Soc. Trans. 31, 20-24.
Application References
- Kappas, N.C. et al. (2008) J Cell Biol 181, 847-58. This article references the use of Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb in the following applications: IC-IF Western Blotting
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
- 2904 VEGF Receptor 2 Control Proteins
- 9943 Vascular Endothelial Growth Factor (VEGF)
- 2479 VEGF Receptor 2 (55B11) Rabbit mAb
- 4991 Phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb
- 2477 Phospho-VEGF Receptor 2 (Tyr1212) (11A3) 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.
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.