Flow Cytometry Phenotyping of BM-DCs

BM-DCs were suspended in cold PBS containing 2.5% FBS, washed, and then incubated in a one-step test with the fluorophore-labeled monoclonal antibodies according to the procedure described in previous article [15 (link)] (mAbs): hamster anti-mouse APC-CD80 (BD Pharmingen, USA, clone 16-10A1), rat anti-mouse PE-Cy7-CD86 (BD Pharmingen, USA, clone GL1), mouse anti-mouse FITC I-A^b (BD Pharmingen, USA, clone 25-9-17), rat anti-mouse RPE-CD40 (BD Pharmingen, USA, clone 3/23), and the appropriate isotype controls: APC-labeled Hamster IgG2,k (BD Pharmingen, USA, clone B83-3), PE-Cy7-labeled Rat IgG2a (BD Pharmingen, USA, clone R35-95), FITC-labeled Mouse IgG2a (BD Pharmingen, USA, clone G155-178), R-phycoerythrin (RPE)-labeled IgG2a (BD Pharmingen, USA, clone R35-95). The cells were stained for 45 min at 4°C. The analysis was carried out using Becton Dickinson FACSCalibur apparatus.