Apc cy7 conjugated cd19

Single cell suspensions of splenocytes were resuspended in PBS containing 2% fetal bovine serum and stained on ice for 20 minutes. Prior to staining, Fc receptors were blocked with anti-16/32 (BD Biosciences). Antibodies used were BV421 conjugated anti-Igλ (BD Biosciences), PE conjugated anti- Igκ (BD Biosciences), PE-Cy7 conjugated anti-CD95 (BD Biosciences), APC-Cy7 conjugated CD19 (BD Biosciences), PE conjugated CD138 (BD Biosciences), PerCP-Cy5.5 conjugated anti-CD3, CD4 and CD8 (Biolegend), Pac Blue conjugated anti-B220, and E-Fluor 660 conjugated anti-GL7 (eBioscience).
For each infection, a separate group of C57Bl6/J mice were infected in parallel with wild type MHV68. Gates for YFP+ populations were set based on the level of auto-fluorescence detected in the FITC channel from mice infected with wt virus. Single cells were sorted based on YFP and surface marker expression (Table 1). Cells were sorted into 96 well plates containing 4 ul of ice cold 0.5X PBS supplemented with 8U/ul of RNAsin (Promega). After sorting, plates were snap frozen in a dry ice/ethanol bath and stored at -80⁰C.