Multicolor Flow Cytometry of Murine Blood and Bone Marrow

For multicolor flow cytometry analysis whole blood and bone marrow (BM) of the mice was collected. Erythrocytes in 100µl whole blood were lysed using a TQ-Prep Workstation (Beckmann Coulter, Brea, CA, USA), for analysis of BM, 1x10⁵ cells/stain were used. Cells (lysed blood cells and BM) were resuspended in 50µl Fc-block solution (eBioscience, San Diego, CA, USA) and incubated for 10min at room temperature. Staining for antibody panels was then carried out at 4°C for 30min as described previously (39 (link)). Cells were analyzed using a CytoFLEX S flow cytometer and data was analyzed with the help of the Kaluza analysis software (both: Beckman Coulter, Brea, CA, USA).