Miltenyi, 130-094-775, CD4+CD25+CD127dim/- Regulatory T Cell Isolation Kit II, human

Background information
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.

Detailed separation procedure
First, the isolation of CD4+CD25+CD127dim/– regulatory T cells was performed with a cocktail of biotinylated antibodies and Anti-Biotin MicroBeads for the depletion of non-CD4+ and CD127high cells. In the second step, the flow-through fraction of pre-enriched CD4+CD127dim/– T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4+CD25+CD127dim/– regulatory T cells.

Downstream applications
Isolation of CD4+CD25+CD127dim/– regulatory T cells from human PBMCs for further phenotypical or functional characterization.

Columns
For the first magnetic separation (depletion): LD or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

Overview
The CD4+CD25+CD127dim/– Regulatory T Cell Isolation Kit II was developed for the isolation of CD4+CD25+CD127dim/– Treg cells from human PBMCs. The isolation is performed in a two-step procedure.