YILIMART - Memory B Cell Isolation Kit, mouse

The Memory B Cell Isolation Kit was developed for the isolation of memory B cells from mouse spleen and lymph nodes.

Background information

Memory B cells are quiescent B cells derived from proliferating antigen-experienced precursors. They are able to quickly react to a recurrent antigenic challenge, thereby providing humoral immune protection.

Detailed separation procedure

The isolation of mouse memory B cells is performed in two steps: First, non-memory B cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies, as primary labeling reagent and subsequently depleted using Anti-Biotin MicroBeads. During the first incubation step with the cocktail, cells are additionally labeled with Anti-IgG1-APC and Anti-IgG2ab-APC antibodies. In the second step, pre-enriched memory B cells are labeled with Anti-APC MicroBeads and isolated by positive selection over a MACS Column.

Columns

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

Figure 1

Memory B cells were isolated from mouse spleen using the Memory B Cell Isolation Kit, mouse. Cells were fluorescently stained with CD45R (B220)-VioBlue (#130-094-287), CD38-PE (#130-097-087) and Anti-IgM-FITC (#130-095-906) and analyzed by flow cytometry using the MACSQuant

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Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Unseparated fraction


After separation

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Memory B Cell Isolation Kit, mouse

For research use only

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Background information

Detailed separation procedure

Columns

Figure 1

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