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| Regulatory B Cell Isolation Kit |
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| Overview |
| The Regulatory B Cell Isolation Kit was developed for the isolation of viable IL-10-producing regulatory B cells from single-cell suspensions of lymphoid organs. |
| Details |
Detailed procedure
The isolation of regulatory B cells is performed in three steps. First, B cells are pre-enriched by depletion of non-B cells. Second, the pre-enriched B cells are stimulated for 5 hours in culture with, e.g., LPS, PMA, and ionomycin to induce IL-10 secretion. Third, the viable IL-10–producing cells are specifically isolated by using the Cytokine Secretion Assay technology.
Downstream applications
Isolated viable IL-10 secreting regulatory B cells can be used for phenotyping and functional analysis. |
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| Regulatory B cells were isolated from mouse spleen using the Regulatory B Cell Biotin-Antibody Cocktail, Anti-Biotin MicroBeads, an LD Column, and a MidiMACS™ Separator. Cells were fluorescently stained with CD19-APC (#130-092-039) and CD4-FITC (#130-091-608) and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. |
| Pre-enriched B cells |
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| The pre-enriched regulatory B cells were incubated for 5 hr with and without LPS, PMA, and ionomycin. The isolation of IL-10-producing B cells was performed by using the principle of the Cytokine Secretion Assay. Analysis of IL-10 (PE) versus CD19-APC staining of viable lymphocytes is displayed. |
| Stimulated sample |
| Before enrichment |
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| After enrichment |
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| Unstimulated control sample |
| Before enrichment |
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| After enrichment |
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| Favorites / Prices |
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| Details |
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| Products |
| Regulatory B Cell Isolation Kit, mouse |
- for 2x109 total cells
Download datasheet
130-095-873
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