Rabbit anti von willebrand factor antibody

Particle size distributions were measured by DynaPro Plate Reader II (Wyatt Technology Corporation, Santa Barbara, CA), in three replicates, 5 acquisitions of 5 s each at 25°C. PBS buffer and UC-isolated cell line EV were used as controld; direct CD9 ELISA was performed as follows: 1 μg of the protein from neat plasma, plasma with cell line EV (Vcap) precipitated by F68 and plasma alone precipitated by F68 were coated on the 96-well ELISA plate overnight at 4°C, the plates were blocked by 1% BSA in PBS for 1 h; biotinylated anti-CD9 monoclonal antibody (Abcam, ab34161) was added as detection antibody followed by streptavidin-HRP; 100 μL of Ultra ELISA Substrate (Pierce) was used and OD450 was recorded by the BioTek plate reader; for western blot, protein samples were separated on 4–12% Bis Tris protein gel and transferred to a polyvinylidene difluoride (PDVF) membrane. Primary antibody includes mouse anti-CD9 (Abcam), mouse anti-β actin antibody (Abcam), mouse anti-HSP70 antibody (Abcam), rabbit anti-Von Willebrand Factor antibody (Abcam), rabbit anti-human serum albumin antibody (Abcam), mouse anti-alpha2 macroglobulin antibody (Abcam) and rabbit anti-human IgG antibody (Abcam). Chemiluminescent signals were captured by imager (PXi, Syngene).