CD4+CD25+CD45RA+ Regulatory T Cell Isolation Kit
Description The CD4+CD25+CD45RA+ Regulatory T Cell Isolation Kit has been developed for the isolation of CD45RA+ regulatory T cells from human PBMCs. The isolation is performed in a two-step procedure. First, non-CD4+ and memory T cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted by separation over a MACS® Column. In the second step, CD4+CD25+CD45RA+ T cells are directly labeled with CD25 MicroBeads and isolated by positive selection from the pre-enriched CD4+CD45RA+ T cell fraction. Regulatory CD4+CD25+ T cells isolated from the CD45RA+ naive T cell compartment were shown to be optimal for in vitro expansion. These expanded regulatory T cells maintained the FoxP3+ phenotype and their suppressive activity, whereas CD45RA– regulatory T cells from the memory T cell compartment lost FoxP3 expression and showed only modest suppressive activity.1 Applications Isolation of CD4+CD25+CD45RA+ regulatory T cells from human PBMCs, for example, for further phenotypical or functional characterization and expansion. Columns For depletion of non-CD4+ and memory T cells: LD or autoMACS® Columns. For positive selection of CD25+ cells: MS or autoMACS Columns.		CD4+CD25+CD45RA+ regulatory T cells were isolated from human PBMCs by using the CD4+CD25+CD45RA+ Regulatory T Cell Isolation Kit, an LD, and two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. The isolated cells were fluorescently stained with CD4, CD25, and either CD45RA or Anti-FoxP3 antibodies. Dot plots show data gated on viable lymphocytes (C) or viable CD4+ lymphocytes (A, B). A B C

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