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.

Data and images for Regulatory B Cell Isolation Kit, mouse

Figures

Figure 1

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
View details

Figure 1

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 regulatory B cells after incubation with LPS, PMA, and ionomycin
Before enrichment
After enrichment
View details

Figure 1

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.
View details

Figure 1

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.
Unstimulated control
Before enrichment
After enrichment
View details

Figure 1

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.
View details

Figure 1

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.

Specifications for Regulatory B Cell Isolation Kit, mouse

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.

Detailed product information

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.

Columns

For the first magnetic separation (depletion): LD or autoMACS
®
Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

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Regulatory B Cell Isolation Kit, mouse

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