CD4+CD25+CD127dim/– Regulatory T Cell Isolation Kit II
Description The isolation of CD4+CD25+CD127dim/– regulatory T cells is performed with a cocktail of biotinylated antibodies and Anti-Biotin MicroBeads for the depletion of non-CD4+ and CD127high cells, and CD25 MicroBeads II for the subsequent positive selection of CD4+CD25+CD127dim/– cells. CD127, the α-chain of the IL-7 receptor, is expressed on the majority of mature T cells and plays an important role in their proliferation and differentiation. However, on regulatory T cells CD127 is absent and its expression inversely correlates with FoxP3 expression. Thus, CD127 can be used as an additional marker to discriminate between human regulatory and activated T cells. Applications Isolation of CD4+CD25+CD127dim/– regulatory T cells from human PBMCs for further phenotypical or functional characterization Columns For depletion of non-CD4+ and CD127high cells: LD or autoMACS® Columns. For positive selection of CD25+ cells: MS or autoMACS Columns.		Figure 1 CD4+CD25+CD127dim/– regulatory T cells were isolated from human PBMCs by using the CD4+CD25+CD127dim/– Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot. Before isolation A B Isolated CD4+CD25+CD127dim/- regulatory T cells A B C Pre-enriched CD4+CD127dim/- cells A

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