Anti cd14 apc eflour780

Heparinized whole blood was drawn from all study participants. Blood was diluted 1:1 (v:v) with RPMI 1640 (Sigma–Aldrich), added anti‐C3aR‐AF647 (BD), anti‐CD3‐BV711 (BD), anti‐CD14‐APC‐eflour780 (ebioscience), anti‐CD88‐PE‐Cy5 (Biolegend), anti‐CD123‐BV650 (BD), anti‐CD124‐AF700 (R&D) anti‐CD193‐BV421 (BD) and anti‐CD294‐PE‐CF594 (BD) or stained with anti‐CD3‐BV711 (BD), anti‐CD14‐APC‐eflour780 (ebioscience), anti‐CD32‐FITC (BD), anti‐CD123‐BV650 (BD), anti‐CD172a‐PerCP‐eFlour^®710 (ebioscience), anti‐CD193‐BV421 (BD), anti‐CD200R‐PE (Biolegend), anti‐CD300a‐AF647 (Novus) and anti‐Siglec‐8‐AF700 (R&D Systems). Blood was incubated with or without 1 μg/ml polyclonal goat anti‐human IgE (KPL) for 30 min at 37°C using a water bath. Erythrocytes were lysed with BD FACS™ lysing solution (BD). Cells were washed and stained with anti‐FcϵRI‐PE‐Cy7 (Biolegend) for 30 min at 4°C. Cells were washed, fixated using BD CellFix™ (BD) and acquired on a BD Fortessa flow cytometer. Data were analyzed as described above. Basophils were gated as CD3^‐CD14^‐CD193⁺CD123⁺ cells (Fig. S2A).