描述：

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.