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| CD4+CD25+CD127dim/– Regulatory T Cell Isolation Kit II |
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| Description |
| The CD4+CD25+CD127dim/– Regulatory T Cells Isolation Kit II was developed for the isolation of CD4+CD25+CD127dim/– Treg cells from human PBMCs. The isolation is performed in a two-step procedure. |
| Details |
Background information
CD127, the α-chain of the IL-7 receptor, is expressed on the majority of mature T cells and plays an important role in their proliferation and differentiation. However, on regulatory T cells CD127 is absent and its expression inversely correlates with FoxP3 expression. Thus, CD127 can be used as an additional marker to discriminate between human regulatory and activated T cells.
Detailed separation procedure
First, the isolation of CD4+CD25+CD127dim/– regulatory T cells was performed with a cocktail of biotinylated antibodies and Anti-Biotin MicroBeads for the depletion of non-CD4+ and CD127high cells. In the second step, the flow-through fraction of pre-enriched CD4+CD127dim/– T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4+CD25+CD127dim/– regulatory T cells.
Downstream 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 II, 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, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot. |
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| Isolated CD4+CD25+CD127dim/- regulatory T cells |
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| Pre-enriched CD4+CD127dim/- cells |
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| Details |
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| Products |
| CD4+CD25+CD127dim/– Regulatory T Cell Isolation Kit II, human |
- for 109 total cells
Download datasheet
130-094-775
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| References |
| 1. Seddiki et al. (2006) J. Exp. Med. 203: 1693–1700. |
| 2. Liu et al. (2006) J. Exp. Med. 203: 1701–1711. |
| 3. Stockis et al. (2009) Eur. J. Immunol. 39: 3315–3322. |
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