Vascular Endothelial Growth Factor Receptor 2 (VEGFR2) also known as kinase insert domain receptor is a VEGF receptor. There are three main subtypes of VEGFR, numbered 1, 2 and 3. All members of the VEGF family stimulate cellular responses by binding to VEGF receptors on the cell surface, causing them to dimerize and become activated through transphosphorylation. VEGFR2 appears to mediate almost all of the known cellular responses to VEGF. Besides, Vascular Endothelial Growth Factor C (VEGFC) has been identified as an interactor of VEGFR2, thus a binding ELISA assay was conducted to detect the interaction of recombinant human VEGFR2 and recombinant human VEGFC. Briefly, VEGFR2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to VEGFC-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-VEGFR2 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°C. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of VEGFR2 and VEGFC was shown in Figure 1, and this effect was in a dose dependent manner.

(A) ECV-304 cells cultured in DMEM, stimulated with 10ng/mL VEGFR2 for 96h; (B) Unstimulated ECV-304 cells cultured in DMEM for 96h.