| Description |
The CD4+CD25+CD127dim/- Regulatory T Cell Isolation Kit has been developed for the isolation of CD4+CD25+CD127dim/- cells from PBMCs. The kit contains a cocktail of biotinylated antibodies and Anti-Biotin MicroBeads for the depletion of non-CD4+ and CD127high cells, and CD25 MicroBeads for the subsequent positive selection of CD4+CD25+CD127dim/- cells.
Regulatory CD4+CD25+ T cells suppress harmful immunological reactions to self or foreign antigens and act upon T cell receptor–mediated activation.
CD127, the α-chain of the IL-7 receptor, is expressed on most mature T cells and plays an important role in their differentiation and proliferation. CD127 expression is down-regulated on human regulatory CD4+ T cells and its expression is inversely correlated with FoxP3 expression. Thus, CD127 can be used as an additional marker to distinguish between human regulatory T cells and conventional T cells. |
| Applications |
| Isolation of CD4+CD25+CD127dim/- regulatory T cells from human PBMCs for further phenotypical or functional characterization. |
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| Figure 1 |
| CD4+CD25+CD127dim/- regulatory T cells were isolated from human PBMCs by using the CD4+CD25+CD127dim/- Regulatory T Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™Separator and a MiniMACS™Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE (A), or CD4-FITC, CD127-PE, and Anti-FoxP3-APC (B). Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot. |
| A) |
| Before enrichment |
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| Pre-enriched CD4+ T cells after depletion of non-CD4+ and CD127high cells |
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| Isolated CD4+CD25+CD127dim/- regulatory T cells |
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| Isolated CD4+CD25+CD127dim/- regulatory T cells |
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| B) |
| Before enrichment |
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| Isolated CD4+CD25+CD127dim/- regulatory T cells |
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