CD4+CD25+CD45RA+ Regulatory T Cell Isolation Kit ( #130-093-631 )

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.

Figure

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).