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 - Excellent tool to isolate both Treg cells and CD4+ T cells for functional analysis
- Cells can be directly used for downstream assays
- Outstanding purity and unmatched recovery
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| Description |
| The CD4+CD25+ Regulatory T Cell Isolation Kit was developed for the isolation of CD4+CD25+ cells from human PBMCs. The isolation is performed in a two-step procedure. |
| Details |
Background information
Regulatory CD4+CD25+ T cells are suppressor cells that neutralize other immune cells by various mechanisms. Their characteristic marker is the transcription factor FoxP3. Regulatory CD4+CD25+ T cells seem to suppress harmful immunological reactions to self or foreign antigens.
Detailed separation procedure
The isolation is performed in a two-step procedure. First, non-CD4+ T cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies against CD8, CD14, CD16, CD19, CD36, CD56, CD123, TCRγ/δ and CD235a (glycophrin A) 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 CD4+CD45RA+ T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4+CD25+ regulatory T cells.
Downstream applications
The CD4+CD25+ Regulatory T Cell Isolation Kit has been used to investigate the role of CD4+CD25+ regulatory T cells in autoimmune diseases1,2, antitumor immunity3–6, allergy7, and in infectious diseases8. For further characterization, CD4+CD25+ regulatory T cells were isolated from patients suffering from rheumatoid arthritis1 or multiple sclerosis2, from acute T cell lymphoma patients4, from tumor biopsy specimens of non-Hodgkin lymphoma patients5, after dendritic cell vaccination9 as well as from patients with atopic dermatitis7. |
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| Figure 1 |
| CD4+CD25+ regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot. |
| Before separation |
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| Isolated CD4+CD25+ regulatory T cells |
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| Pre-enriched CD4+ T cells |
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| Details |
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| Products |
| CD4+CD25+ Regulatory T Cell Isolation Kit, human |
- for 109 total cells
Download datasheet
130-091-301
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| References |
| 1. Ehrenstein et al. (2004) J. Exp. Med. 200: 277-285. |
| 2. Hong et al. (2006) Proc. Natl. Acad. Sci. USA 103: 5024-5029. |
| 3. Nishikawa et al. (2006) J. Immunol. 176: 6340-6346. |
| 4. Karube et al. (2004) Br. J. Haematol. 126: 81-84. |
| 5. Yang et al. (2006) Blood 107: 3639-3646. |
| 6. Mougiakakos et al. (2009) Blood 113: 3542-3545. |
| 7. Ou et al. (2004) J. Allergy Clin. Immunol. 113: 756-763. |
| 8. Xu et al. (2006) J. Immunol. 177: 739-747. |
| 9. Schwaab et al. (2009) Clin. Cancer Res. 15: 4986-4992. |
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