Active Vascular Endothelial Growth Factor 165 (VEGF165)

Figure. The binding activity of VEGF165 with VEGFR1.
Vascular endothelial growth factor 165 is one kind of isoforms of Vascular endothelial growth factor A (VEGFA). This protein is a glycosylated mitogen that specifically acts on endothelial cells and has various effects, including mediating increased vascular permeability, inducing angiogenesis, vasculogenesis and endothelial cell growth, promoting cell migration, and inhibiting apoptosis. Besides, Vascular Endothelial Growth Factor Receptor 1 (VEGFR1) has been identified as an interactor of VEGF165, thus a binding ELISA assay was conducted to detect the interaction of recombinant human VEGF165 and recombinant human VEGFR1. Briefly, VEGF165 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to VEGFR1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-VEGF165 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of VEGF165 and VEGFR1 was shown in Figure 1, and this effect was in a dose dependent manner.