CD4+ Recent Thymic Emigrant Isolation Kit, human130-094-299
产品说明
Background information
RTEs are a subpopulation of naive T cells characterized by their content of T cell receptor excision circles (TRECs).
CD31 (platelet endothelial cell adhesion molecule-1, PECAM-1) has been described as a marker to identify the CD4+ RTEs among the naive CD4+ T cell population, as its expression correlates with the TREC content of CD4+ T cells1. CD4+ T cells lacking CD31 expression harbor little or no TRECs, suggesting that they already proliferated in the periphery.
Therefore, the magnetic isolation of CD4+ RTEs is possible by direct isolation of CD31+ cells from pre-enriched, untouched naive CD4+ T cells.
Detailed separation procedure
The isolation is performed in a two-step procedure. First, non-CD4+ and memory CD4+ T cells are indirectly magnetically labeled with a cocktail of biotinylated antibodies 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 naive CD4+ T cells is labeled with CD31 MicroBeads for subsequent positive selection of CD4+CD45RA+CD31+ RTEs.
Columns
For the first magnetic separation (depletion): LS or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.
相关图片
Isolation of CD4+ RTEs from PBMCs using the CD4+ Recent Thymic Emigrant Isolation Kit, an LS Column, and an MS Column. To evaluate the isolation of CD4+ RTEs, PBMCs before separation and the isolated CD4+ RTEs were fluorescently stained with CD4, CD45RA, and CD31 antibodies and analyzed by flow cytometry.
PBMCs before separation
PBMCs before separation gated on CD4+CD45RA+cells
Isolated CD4+ RTEs
CD4+ RTEs gated on CD4+CD45RA+ cells