Mouse anti human igg2b cd14 apc

10% of a FcR blocking reagent (Myltenyi Biotec) was added and incubated for 10- 30 minutes on ice. 100 μl of staining buffer was added per 10⁷ of cells. The PBMCs were then stained with the following antibodies: mouse anti-human IgG2b CD14 –APC and mouse anti-human IgG1 CD16-PE (BD bioscience) and incubated for 30–45 minutes on ice in the dark. Finally, the cells were spun at 200 × g for 15 minutes and the pelleted PBMCs were resuspended in 1 mL of staining buffer for sorting. Fluorescence (FL) was quantified on a SORP FACSAria2 (BD Biosciences). Data were processed with FACSDiva 6.3 software (BD Biosciences). Doublets were excluded by FSC-W × FSC-H and SSC-W × SSC-H analysis (Supplementary Figure 4). PE fluorescent was acquired by 498 nm blue laser and 575/26 nm emission; APC fluorescence was obtained by 650 nm red laser and 660/20 nm emission; Figures display the median of fluorescence intensity (mfi) relative to control. Single stained channels were used for compensation and fluorescence minus one (FMO) controls were used for gating. Purity of the sorting was controlled after each sorting and was > 90%.