CD25+CD49d– Regulatory T Cell Isolation Kit
Description 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. 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. Applications Isolation of CD4+CD25+ regulatory T cells from PBMCs for further phenotypical or functional characterization. Columns For depletion of CD49d+ cells: LD or autoMACS® Columns. For further concentration of CD25+: MS or autoMACS® Columns.		CD25+CD49d– regulatory T cells were isolated from human PBMCs using the CD25+CD49d– Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. Before isolation A B Pre-enriched CD25+CD49d– Treg cells A B Isolated CD25+CD49d– Treg cells A B C: Gated on CD4+ T cells

Search this website

Advanced search