Apc rat igg

Dissociated prostate cells were suspended in DME-10% FBS, preincubated with purified rat anti-mouse CD16/CD32 (mouse BD Fc Block) antibody (1:50, BD Pharmingen, San Jose, CA) at 4°C for 10 min, and stained with specific antibodies at 4°C for 30 min. Flow cytometry analysis was performed using a BD FACS Canto II and analysed by NovoExpress^™ software (ACEA Biosciences, San Diego, CA). Cell sorting was conducted on a BD FACS Vantage or a BD FACS Aria. Primary antibodies used were: PE-Cy5 rat anti-human/mouse CD49f (1:100, BD Pharmingen), APC rat anti-human/mouse CD49f (1:100, BD Pharmingen), APC rat anti-mouse CD24 (1:100, BD Pharmingen), PE rat anti-mouse Sca-1 (1:100, eBioscience, San Diego, CA). Isotype controls used were from BD Pharmingen : PE rat IgG (1:100), PE-Cy5 Rat IgG (1:100), APC Rat IgG (1 :100). For lineage staining, biotinylated antibodies used were : rat anti-mouse CD31 (1:100, BD Pharmingen), rat anti-mouse CD45 (1:100, eBioscience), rat anti-mouse TER-119 (1:100, eBioscience). Dynabeads M-280 Streptavidin (Invitrogen) or BD Horizon™ V450-Streptavidin (1 :10000, BD Biosciences, Franklin Lakes, NJ) were used.