Product Pathways - Tyrosine Kinase / Adaptors
Phospho-VEGF Receptor 2 Antibody Sampler Kit #12599
|12599S||1 Kit (5 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb #4991||40 µl||W, IHC-P||H, M||230||Rabbit IgG|
|Phospho-VEGF Receptor 2 (Tyr996) Antibody #2474||40 µl||W||H, M||230||Rabbit|
|Phospho-VEGF Receptor 2 (Tyr1059) (D5A6) Rabbit mAb #3817||40 µl||W||H, M||R||230||Rabbit IgG|
|Phospho-VEGF Receptor 2 (Tyr1175) (D5B11) Rabbit mAb #3770||40 µl||W, IP||H, M||230||Rabbit IgG|
|VEGF Receptor 2 (D5B1) Rabbit mAb #9698||40 µl||W, IP, IHC-P, IF-F, F||H, M, R||210, 230||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IP=Immunoprecipitation, IF-F=Immunofluorescence (Frozen), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat
Western blot analysis of extracts from PAEC/CKR cells expressing chimeric receptors containing human CSF-1 extracellular binding domain/mouse VEGF receptor 2 intracellular domains (Rahimi, N. et al. (2000) J. Biol. Chem. 275, 16986-16992), using Phospho-VEGF Receptor 2 (Tyr996) Antibody #2474 (upper) or VEGF receptor 2 antibody (lower).
Western blot analysis of extracts from HUVEC, untreated (-) or stimulated with vascular endothelial growth factor (VEGF; +), using Phospho-VEGF Receptor 2 (Tyr1175) (D5B11) Rabbit mAb #3770 (upper) and VEGF Receptor 2 (55B11) Rabbit mAb (lower) #2479.
Western blot analysis of PAEC/CKR cells, untreated (-) or stimulated with CSF-1 (+), using Phospho-VEGF Receptor 2 (Tyr1059) (D5A6) Rabbit mAb #3817 (upper) and VEGF Receptor 2 (55B11) Rabbit mAb #2479 (lower). PAEC/CKR cells express a chimeric receptor made up of human CSF-1 receptor extracellular domain and mouse VEGF receptor 2 transmembrane and intracellular domains.
Phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb #4991 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 #4991 (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 #4991.
The Phospho-VEGF Receptor 2 Antibody Sampler Kit provides an economical means of evaluating the VEGFR2 tyrosine kinase and several phosphorylation sites that are involved in its activation. The kit includes enough antibody to perform four western blot experiments with each primary antibody.
Specificity / Sensitivity
The phospho-VEGF Receptor 2 (Tyr1175) (D5B11) Rabbit mAb detects endogenous VEGF Receptor 2 at tyrosine 1175 and may cross-react with other tyrosine-phosphorylated receptor tyrosine kinases, including VEGFR1 and FLT3. The phospho-VEGF Receptor 2 (Tyr1059) (D5A6) Rabbit mAb
detects endogenous levels of VEGF Receptor 2 at tyrosine 1059 and can detect VEGF receptors 1 and 3 when phosphorylated at corresponding tyrosine residues due to identical sequences within the epitope region. The phospho-VEGF Receptor 2 (Tyr951) (15D2) Rabbit mAb detects endogenous VEGF Receptor 2 at tyrosine 951 and may slightly cross-react with activated VEGF receptor 1, but not with other related tyrosine phosphorylated tyrosine kinases. The phospho-VEGF Receptor 2 (Tyr996) Antibody detects transfected VEGF Receptor 2 only when phosphorylated at tyrosine 996. The antibody cross-reacts with tyrosine-phosphorylated PDGF receptor. The VEGF Receptor 2 (D5B1) Rabbit mAb detects endogenous total VEGF Receptor 2.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr996 of human VEGFR2. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr951, Tyr1059 or Tyr1175. VEGFR2 monoclonal antibody is produced by immunizing animals with a recombinant protein containing the carboxy-terminal 150 residues of human VEGFR2.
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).
- Meyer, M. et al. (1999) EMBO J 18, 363-74.
- Dougher-Vermazen, M. et al. (1994) Biochem Biophys Res Commun 205, 728-38.
- Kroll, J. and Waltenberger, J. (1997) J Biol Chem 272, 32521-7.
- Takahashi, T. et al. (2001) EMBO J 20, 2768-78.
- Holmqvist, K. et al. (2004) J Biol Chem 279, 22267-75.
- Karkkainen, M.J. and Petrova, T.V. (2000) Oncogene 19, 5598-605.
- Rahimi, N. et al. (2000) J Biol Chem 275, 16986-92.
- Claesson-Welsh, L. (2003) Biochem Soc Trans 31, 20-4.
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- 9998 BSA
- 9999 Nonfat Dry Milk
- 9997 Tris Buffered Saline with Tween® 20 (TBST-10X)
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
Tween® is a registered trademark of ICI Americas, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.