Reagents 1×PBS/2%FCS (should be sterile if necessary)
ACK buffer (41.45g NH4Cl, 5g KHCO3, 0.18g EDTA, H2O to 5 liters, store at 4°C, filter to sterilize, if necessary).
Propidium iodide (PI, Sigma), make 500× stock solution (1.0mg/ml) in a fume hood.
Antibodies (The amount of the antibodies provided in this protocol is just an example. It is necessary to titrate all the antibodies every time when using a new batch even if it has the same catalog number.)
Tricolor (TC=phycoerythrin PE-Cy5) conjugated rat antibodies specific for CD3, CD4, CD8, CD19, B220 and Gr-1 (Caltag).
Allophycocyanin (APC)-conjugated anti-c-kit (BD Pharmingen).
FITC-conjugated anti-CD34 (BD Pharmingen).
Biotinylated anti-Sca-1 (BD Pharmingen).
PE-conjugated anti-FcγRIII/II (eBioscience).
Anti B220 antibody conjugated to FITC, PE, APC (BD Pharmingen), TC and APC-Cy7 (Caltag).
RNeasy Micro kit (Qiagen).
β-mercaptoethanol (β-ME, Sigma).