anti-CD14-BV570 (301831, Biolegend), anti-CD11c-PerCP-Cy5.5 (337210), anti-HLA-DR-AlexaFlour700 (307626) all from Biolegend, anti-CD66a/c/e-QDot605 (342302, Biolegend, with in-house conjugation to QDot605, Q-22001), anti-CD16-PE-Cy5 (555408, from BD Biosciences), anti-IL-12-PE (554575), anti-TNF-α-PE-Cy7 (25-7349-82) both from eBioscience, anti-IL-10-PE (506804), anti-IL-6-APC (501112), anti-CD3-BV421 (300434), anti-CD19-BV421 (302233) and anti-CD335-BV421 (331913) all from Biolegend.
Cryopreserved, fixed white cells were washed in PBS and stained for 1 hour at 4°C with surface marker antibodies. For intracellular staining, cells were permeabilized with Perm/Wash buffer (BD Biosciences) and incubated for 1 hour at 4°C with antibodies for intracellular markers. Fixed cells were then acquired on a LSR II flow cytometer (BD Biosciences).
Cryopreserved, fixed white cells were washed in PBS and stained for 1 hour at 4°C with surface marker antibodies. For intracellular staining, cells were permeabilized with Perm/Wash buffer (BD Biosciences) and incubated for 1 hour at 4°C with antibodies for intracellular markers. Fixed cells were then acquired on a LSR II flow cytometer (BD Biosciences).