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T cells were enriched using the CD154 Enrichment and Detection Kit, two MS Columns, and a MiniMACS™ Separator. Spleen cells from a KLH immunized BALB/c mouse were restimulated in vitro with 100 μg/mL KLH or were not restimulated in the presence of the CD154 Detection Cocktail (PE) for 6 hours.
T cells were magnetically isolated using Anti-PE MicroBeads UltraPure. Cells were fluorescently stained with CD4-FITC and CD45R (B220)-PerCP. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide (PI) fluorescence. CD45R+
cells were excluded from the analysis.
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