Anti cd11c allophycocyanin apc

For isolation of splenic dendritic cells and macrophages, mouse spleens were perfused with 400 U/ml of collagenase D (Roche, Basel, Switzerland) in Hanks' balanced salt solution and incubated for 45 min at 37° followed by mechanical dissociation. Splenocytes were first incubated with anti‐CD16/32 (Fc‐block) (Biolegend, San Diego, CA) in PBS [1 mm EDTA, 2% fetal calf serum (FCS)] at 4° for 15 min, and were then stained with anti‐CD11c‐allophycocyanin (APC) (BD Biosciences, San Jose, CA) or anti‐F4/80‐APC (BD Biosciences, San Jose, CA) at 4° in PBS with 1 mm EDTA, 2% FCS. The BMDMs were first incubated with anti‐CD16/32 (Fc‐block) (Biolegend, San Diego, CA) in PBS (1 mm EDTA, 2% FCS) at 4° for 15 min. Cells were then stained with the following panel for 30 min at 4° in PBS (1 mm EDTA, 2% FCS): TLR2‐APC (Biolegend, San Diego, CA), CD206‐phycoerythrin/Cy7 (Biolegend, San Diego, CA), CD38‐BV510 (BD Biosciences, San Jose, CA) and F4/80‐APC/Cy7 (Biolegend, San Diego, CA). After washing twice, the cells were acquired using a Gallios flow cytometer (Beckman Coulter, Brea, CA) followed by data analysis using flowjo v10 (FlowJo, Ashland, OR).