Fitc conjugated cd24 antibody

Prostate tissues were digested in DMEM/F12/Collagenase/Hyaluronidase/FBS (Invitrogen, Carlsbad, CA) for 3 hours at 37°C, followed by an additional 1 hour of digestion in 0.25% Trypsin-EDTA (Invitrogen, Carlsbad, CA) on ice. Subsequently, digested cells were suspended in Dispase (Invitrogen, Carlsbad, CA, 5 mg/mL) and DNase I (Roche Applied Science, Indianapolis, IN, 1 mg/mL), and pipetted vigorously to dissociate cell clumps. Dissociated cells were then passed through 70 µm cell strainers (BD Biosciences, San Jose, CA) to get single cells. For FACS analysis and sorting, singled murine prostate cells were stained with Pacific blue-conjugated anti CD31, CD45, and Ter119 antibodies (eBioscience, San Diego, CA), PE-Cy7-conjugated anti Sca-1 antibody (eBioscience, San Diego, CA), FITC-conjugated CD24 antibody (BD Biosciences, San Jose, CA), and Alexa 647-conjugated anti CD49f antibody (Biolegend, San Diego, CA). Cell sorting and analyses were performed using Aria II and LSR II, respectively (BD Biosciences, San Jose, CA).