1. Pipet 50 pL of whole blood into each of (three) 5-mL tubes (see Note 5). Add 20 pL of the HLA-DR/CD14 MAb mixture to tube 1 (see Note 6). Add 10 pL of CD13 MAb to tube 2 and 10 pL of CD10 MAb to tube 3.
2. Incubate for 30 min in the dark at 4°C (see Note 7).
3. Add 2 mL of FACS Lyse (see Note 8), mix, and incubate for 30 min in the dark at 4°C.
4. Centrifuge for 5 min at 300g and decant supernatant (see Note 9).
5. Analyze cells in 0.5 mL PBS. Store at 4°C in the dark until measurement.
Was this article helpful?