Pe conjugated anti cd16

Ficoll-isolated monocytes (500,000 cells) from fresh PB and BM samples (after the 1–2 days preculture) were stained with 1.25 µl allophycocyanin (APC)-conjugated anti-CD14 (BD Biosciences, Franklin Lakes, NJ; Cat. No. 561383), 5 µl phycoerythrin (PE)-conjugated anti-CD16 (BD Biosciences; Cat. No. 555407), and 1.25 µl fluorescein isothiocyanate (FITC)-conjugated anti-CD3 antibodies (Invitrogen; Cat. No. 11-0036) for 20 min at RT. Ficoll-isolated CB monocytes (500,000 cells) were stained with above-mentioned amounts of APC-conjugated anti-CD14, PE-conjugated anti-CD16 and 2.5 µl FITC-conjugated anti-CD45 antibodies (BD Biosciences; Cat. No. 555482) for 20 min at RT. Samples were run on the BD Accuri 6C Plus (BD Biosciences) flow cytometer and analyzed with FlowJo v10.7.1 software (FlowJo LLC.).