α cd11b pe cy7

Blood cell suspensions were prepared as described above and incubated with the FC blocking antibody α-CD16/32 (eBioscience, San Diego, CA; [5 μg/ml]) for 10 min. The cells were subsequently incubated with the following antibody panel for 25 mins. on ice: α-F4/80-AF488 (AbD serotec, Oxford, UK; [0.5 μg/ml]), α-CD115-PE (Biolegend, San Diego, CA; [1 μg/ml]), α-CD11b-PE/Cy7 (Biolegend; [0.5 μg/ml]), α-Ly6C-PerCP/Cy5.5 (BioLegend; [0.5 μg/ml]), α-Ly6G-APC (BioLegend; [1 μg/ml]), α-CD3e-APC (eBioscience; [1 μg/ml]), α-CD19-APC (eBioscience; [1 μg/ml]), α-CD49b-APC (eBioscience; [1 μg/ml]), and DAPI (Sigma; [0.1ug/ml]). Samples were stained concurrently with fluorescence minus one (FMO) antibody panels. Following a series of washes, the cells were resuspended in MACS buffer, and sorted on the BD Influx Cell Sorter (Becton, Dickinson and Company, Franklin Lakes, NJ). FlowJo software (Tree Star Inc., v.9.2, Ashland, OR) was used for data analysis.