GBM cells were detached with 10 mM EDTA and stained with CS56 antibody at 1∶100 dilutions on ice for 30 min. Cells were incubated with fluorescein isothiocyanate (FITC)-conjugated anti-mouse IgM antibody on ice for 30 min. For measuring cell surface PDGFRA expression, cycloheximide-treated cells were detached and immediately fixed with 4% paraformaldehyde for 15 min. Cells were stained with PDGFRA antibody at 1∶200 dilutions. A FITC-conjugated anti-rabbit IgG was used as the secondary antibody. Florescence intensity was analyzed by FACScan cytometer (BD Pharmingen).
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.