CD4+ Recent Thymic Emigrant Isolation Kit, human

CD4
+
Recent Thymic Emigrant Isolation Kit
, human

The CD4
+
Recent Thymic Emigrant Isolation Kit was developed for the isolation of CD4
+
CD45RA
+
CD31
+
recent thymic emigrants (RTE) cells from human PBMCs in a two-step procedure.

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 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.

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.

  • 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.
    PBMCs before separation
    PBMCs before separation gated on CD4
    +
    CD45RA
    +
    cells
    Isolated CD4
    +
    RTEs
    CD4
    +
    RTEs gated on CD4
    +
    CD45RA
    +
    cells

  • Selected references

    1. Kimmig, S. et al. (2002) Two subsets of naive T helper cells with distinct T cell receptor excision circle content in human adult peripheral blood. J. Exp. Med. 195: 789-794
Product options: 1
130-094-299

CD4
+
Recent Thymic Emigrant Isolation Kit
, human

for 1×
10
9
total cells
CHF  1'116.00

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