Product Name: CD4+CD25+ Regulatory T Cell Isolation Kit, mouse

Description:

The CD4+CD25+ Regulatory T Cell Isolation Kit was developed for the isolation of CD4+CD25+ regulatory mouse T cells from single-cell suspensions of spleen and lymph nodes. The isolation is performed in a two-step procedure.

Detailed separation procedure

The isolation is performed in a two-step procedure. First, non-CD4+ are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies against CD8 (Ly-2), CD11b (Mac-1), CD45R (B220), CD49b (DX5), Ter-119 and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted over a MACS® Column. In the second step, the flow-through fraction of pre-enriched CD4+ T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4+CD25+ regulatory T cells.

Downstream applications:

Regulatory T cells isolated from mouse lymph nodes with the CD4+CD25+ Regulatory T Cell Isolation Kit were used in coculture experiments with dendritic cells to study priming of dendritic cells for tolerance induction in vitro and after adoptive transfer of primed dendritic cells in vivo.1 Furthermore, they were isolated and used in various models for adoptive transfer experiments, e.g. in an experimental autoimmune thyroiditis mouse model, and a melanoma mouse model where they prevented tumor immunity. In addition, in vitro suppression assays were performed with isolated 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.