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I have attempted to use abcam’s Active human VEGFD protein fragment (ab125747; lot: GR89044-2) to induce phosphorylation of VEGFR2. The assay procedure used was to incubate human umbilical cord endothelial cells with the VEGFD at increasing concentrations (6.25-100 ng/ml). Phosphorylation of VEGFR2 was measured by immunoblot using an anti-phosho-VEGFR2(Tyr1175) MAb and also using a phosphor-VEGFR2 specific ELISA. I could not detect phosphorylation of VEGFR2 with either method. A positive control and VEGFC (ab155739) showed phosphorylation so I’m satisfied that the assays worked.
Can you indicate if the product (ab125474) is expected to induce phosphorylation of VEGFR2? And if so can you advise of appropriate conditions.
Asked on Sep 27 2013
Unfortunately, we have not tested this product's ability to induce phosphorylation of VEGFR2 and so we do not know whether it is capable of doing so.
We have only tested it's ability to stimulate the proliferation of Human microvascular endothelial cells (HMVECs) and found the ED50 to be 3-4ng/ml.
Elisa Thomas Abcam Scientific Support
回复于 Sep 27 2013