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B cells were isolated from mouse spleen using the Regulatory B Cell Biotin-Antibody Cocktail, Anti-Biotin MicroBeads, an LD Column, and a MidiMACS™ Separator. Cells were fluorescently stained with CD19-APC (# 130-092-039) and CD4-FITC (# 130-091-608) and analyzed by flow cytometry using the MACSQuant®
Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
Pre-enriched B cells
Pre-enriched regulatory B cells after incubation with LPS, PMA, and ionomycin
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