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Technical Data Sheet

VP2921

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VEGFR-2 (Tyr-801)[conserved site], phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
180 kDa

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Western blot image of GST-recombinant human VEGFR-1 (89 kDa), VEGFR-2 (110 kDa), and VEGFR-3 (86 kDa) C-terminal regions. The blots were probed with rabbit polyclonal anti-VEGFR-2 (a.a. 1304-1317), anti-VEGFR-2 (Tyr-801, conserved site), and anti-VEGFR-3 (a.a. 1285-1298).

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Enlarge

Left: Western blot image of HUVEC cells untreated (-) or treated with pervanadate (1 mM) for 30 min. (+). Right: Western blot image of GST-recombinant VEGFR-2 kinase without (-) or with (+) akaline phosphatase treatment. Both sets of blots were probed with rabbit polyclonal anti-VEGFR-2 (a.a. 1304-1317) or anti-VEGFR-2 (Tyr-801).

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Application
Dilution


ELISA
1:2000


ICC
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
Chen, P. et al. (2014) Proc Natl Acad Sci. 111(15):5514 WB: HUVECs
Chen, T.T. et al. (2010) J Cell Biol. 188(4):595. WB: HUVECs + VEGF
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Background
Vascular endothelial growth factor receptor-2 (VEGFR-2/Flk-1/KDR) is the primary receptor for VEGF in endothelial cells. Other VEGFR family members, VEGFR-1 (Flt-1) and VEGFR-3 (Flt-4), can also transduce the intracellular signals of VEGF. However, the role of VEGFR-1 is observed mainly during embryonic angiogenesis and VEGFR-3 signaling may be restricted to specific types of endothelial cells. Major autophosphorylation sites of VEGFR-2 are located in the kinase insert domain (Tyr-951/996) and in the tyrosine kinase catalytic domain (Tyr-1054/1059). Other sites, Tyr-1175 and Tyr-1212 provide docking sites for downstream signaling molecules. Activation of VEGFR-2 also phosphorylates Tyr-801, leading to PI3-kinase-Akt activation and increases in endothelial nitric oxide synthase activity. Phosphorylation of mutliple sites in VEGFR-2 is required for downstream activation of several signaling pathways that control proliferation, chemotaxis, and sprouting during angiogenesis.


Background References
Dougher-Vermazen, M. et al. (1994) Bioch Biophys Res Com. 205:728.
Meyer, M. et al. (1999) EMBO J. 18:363.
Robinson, C.J. & Stringer, S.E. (2001) J. Cell Sci.114:853.
Garcia Blanes, M. et al. (2007) J Biol. Chem. 282(14):10660.
Immunogen
Phospho-VEGFR-2 (Tyr-801) peptide (coupled to carrier protein) corresponding to amino acids surrounding Tyr-801 in human VEGFR-2. This sequence has high homology to the conserved site in rat and mouse VEGFR-2, and has significant homology to the conserved sites in VEGFR-1 (Tyr-794) and VEGFR-3 (Tyr-812).
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100μl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
This antibody was affinity purified with phospho-VEGFR-2 (Tyr-801) peptide. The purified antibody detects a 180 kDa* band corresponding to VEGFR-2 in western blots of human endothelial cells treated with pervanadate, and shows strong reactivity toward recombinant human VEGFR-2. This reactivity is removed by akaline phosphatase treatment. In addition, the antibody detects the conserved site in VEGFR-1 (Tyr-794). The antibody also works for immunofluorescent staining of VEGFR-2 in HUVEC cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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