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Two-color flow cytometric analysis of I-A/I-E MHC class II alloantigen expression on mouse splenocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 553092/561880) and APC Rat Anti-Mouse CD11b (Cat. No. 553312/561690) antibodies, and either BB515 Rat IgG2a, κ Isotype Control (Cat. No. 564418; Left Panel), BD Horizon BB515 Rat Anti-Mouse I-A/I-E (Cat. No. 565254; Middle Panel), or FITC Rat Anti-Mouse I-A/I-E (Cat. No. 553623/562009; Right Panel). Two-color flow cytometric contour plots showing the expression of I-A/I-E MHC class II alloantigens versus CD45R/B220 and CD11b were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Mouse splenic leucocytes from SJL mice (H-2s haplotype) did not stain positively with the 2G9 antibody (data not shown). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.