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Multicolor flow cytometric analysis of mouse IgM expression on mouse splenocytes. 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 stained with PE Hamster Anti-Mouse CD3e (Cat. No. 561824/553063/553064) and either APC Rat IgG2a κ Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse IgM antibody (Cat. No. 550676/560232; Right Plot) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of IgM (or Ig Isotype control) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer Sy553stem and FlowJo™ software.
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