Rabbit monoclonal anti cd9

For western blot (WB) analysis, the purified extracellular vesicles were lysed with radio-immunoprecipitation assay buffer (Santa Cruz Biotechnology, Dallas, TX, USA), and the cleared lysate was collected by centrifugation for protein separation on 12% sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) gels. After electrophoresis, the separated proteins were transferred to 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore, USA). The membranes were blocked for 1 h with Tris-buffered saline containing Tween 20 (TBST) with 5% non-fat milk. The blots were then incubated with primary antibody at 4°C overnight. The primary antibodies used included mouse monoclonal anti-CD63 (Abcam, Cambridge, UK), anti-ASFV p30 (Prepared by our laboratory), rabbit monoclonal anti-CD9 (Abcam, Cam bridge, UK), anti-APOA1(Abcam, Cam bridge, UK), rabbit polyclonal anti-ASFV p72 (prepared by our laboratory), SERPINC1 (Abcam, Cam bridge, UK). After washing three times with TBST, the membranes were incubated with horseradish peroxidase (HRP)-labeled secondary antibody (Proteintech, Chicago, IL, USA) for 2 h at room temperature. Finally, the proteins were visualized with Clarity enhanced chemiluminescence (ECL) WB substrate (Bio-Rad Laboratories, Hercules, CA, USA).