| Product Includes | Product # | Volume | Cap Color | Storage Temp |
|---|---|---|---|---|
| VEGFR-2 Capture Mouse mAb (100X) | 70208 | 400 µl |
|
+4C |
| VEGFR-2 Detection Rabbit mAb (100X) | 88678 | 400 µl |
|
+4C |
| Anti-rabbit IgG, HRP-linked Antibody (1000X) | 25944 | 40 µl |
|
-20C |
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Description
CST's PathScan® Total VEGFR-2 Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total VEGFR-2 Sandwich ELISA Kit #7340. 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 on a 96 well microplate overnight in PBS. After blocking, cell lysates are added followed by a VEGFR-2 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 total VEGFR-2 protein.
Reagents not supplied
Phosphate Buffered Saline with Tween-20 (PBST-20X) #9809
Cell Lysis Buffer (10X) #9803
TMB Substrate #7004
STOP Solution #7002
Blocking Buffer: 1X PBS/0.5% Tween-20, 1% BSA
96 Well Microplates**
Microplate Reader
** Antibody Pairs have been validated on Corning© 96 Well Clear Polystyrene High Bind Stripwell™ Microplates (#2592).
Notes: Antibody pairs have been optimized using recommended buffers, reagents, plates and the included protocol. Solutions should be made fresh daily.
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).
- 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.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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