CD4+ Recent Thymic Emigrant Isolation Kit ( #130-094-299 )

CD4⁺ Recent Thymic Emigrant Isolation Kit

Description

The CD4⁺ Recent Thymic Emigrant Isolation Kit is designed for the isolation of CD4⁺CD45RA⁺CD31⁺ recent thymic emigrants (RTE) from human PBMCs.
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, CD4⁺CD45RA⁺CD31⁺ RTEs are directly labeled with CD31 MicroBeads and isolated by positive selection from the pre-enriched naive CD4⁺ T cell fraction.
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 cells.¹ 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.

Applications

Isolation of CD4⁺CD45RA⁺CD31⁺ recent thymic emigrants from human PBMCs.

Columns

LS, MS and autoMACS™ Columns.

Figure 1

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.

Before separation

Gated on CD4⁺CD45RA⁺ cells

Isolated CD4⁺ RTEs

Gated on CD4⁺CD45RA⁺ cells