武汉云克隆科技股份有限公司

Von Willebrand Factor (vWF) , a large multimeric plasma glycoprotein , is synthesized mainly by endothelial cells and megakaryocytes. The protein consists of multiple subunits, forming high - molecular - weight multimers. These multimers have emerged as a key player in both primary and secondary hemostasis.Besides,Von Willebrand Factor (vWF) specifically interacts with the Integrin Alpha 2B (ITGa2B) of the platelet membrane receptor complex, forming a critical molecular interaction that facilitates platelet aggregation and subsequent thrombus formation, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant mouse vWF and recombinant human ITGa2B . Briefly, biotin-linked vWF were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to ITGa2B-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 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 vWF and ITGa2B was shown in Figure 1, the EC50 for this effect is 1.36µg/mL.