Miltenyi, 130-094-551, CD25+CD49d– Regulatory T Cell Isolation Kit, human

Background information
CD49d is the α-chain of the integrin VLA-4 and is present on the majority of pro-inflammatory effector cells but absent on FoxP3+ cells. Consequently, the depletion of CD49d+ cells is a useful tool to isolate highly pure populations of CD25+CD49d– regulatory T cells.

Detailed separation procedure
The isolation of CD25+CD49d– regulatory T cells is performed in a two-step procedure. First, CD8+ and CD49d+ cells are magnetically labeled with a cocktail of CD8 and CD49d MicroBeads for depletion. In the second step, CD49d– regulatory T cells are labeled with CD25 MicroBeads and isolated by positive selection from the pre-enriched T cell fraction.
The depletion of CD49d+ cells removes contaminating CD25+ effector T cells1, thus providing highly pure populations of regulatory T cells.

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

Overview
The CD25+CD49d- Regulatory T Cell Isolation Kit was developed for the isolation of CD4+CD25+CD49d- regulatory T cells from human PBMCs in a two-step procedure.