YILIMART - B-1a Cell Isolation Kit, mouse

The B-1a Cell Isolation Kit has been developed for the isolation of CD5

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B-1a cells from mouse body cavities or spleen.

Background information

B-1a cells seem to be involved mainly in T cell–independent and innate-like immune responses and are the main producers of natural antibodies. B-1 cells expressing CD5 are known as B-1a cells and those lacking expression of CD5, but having other hallmarks of B-1 cells, are known as B-1b cells.

Detailed separation procedure

The isolation of B-1a cells is performed in a two-step procedure. First, non-B-1a cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies, as primary labeling reagent, and Anti-Biotin MicroBeads, as secondary labeling reagent. In parallel the target cells are stained with CD5-APC as primary labeling reagent. The labeled cells are subsequently depleted by separation over a MACS

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Column. In the second step, the B-1a cells are indirectly labeled with Anti-APC MicroBeads and isolated by positive selection from the pre-enriched B-1a cell fraction by separation over a MACS Column. After removing the column from the magnetic field, the magnetically retained B-1a cells can be eluted as the positively selected cell fraction.

Columns

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

Figure 1

B-1a cells were isolated from mouse spleen (A, B) or peritoneal cavity (C, D) using the B-1a Cell Isolation Kit, mouse. Cells were fluorescently stained with CD19-PE and CD5-APC 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.

Separation of B-1a cells from mouse spleen

A:	B:
Unseparated fraction	After separation

Separation of B-1a cells from peritoneal cavity

C:	D:
Unseparated fraction	After separation

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B-1a Cell Isolation Kit, mouse

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Background information

Detailed separation procedure

Columns

Figure 1

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