Miltenyi, 130-100-366, MZ and FO B Cell Isolation Kit, mouse

Background information
The isolation of CD21hiCD23lo/– marginal zone (MZ) and CD21int CD23hi follicular (FO) B cells is performed in a two-step procedure. First, the non-B cells, the B-1 cells, and the transitional B 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 between the two labeling steps no washing steps are required. The labeled cells are subsequently depleted by separation over a MACS® Column, which is placed in the magnetic field of a MACS Separator. The unlabeled fraction contains the pre-enriched MZ and FO B cells. In order to further separate these two subsets, the FO B cells are directly labeled with CD23 MicroBeads in a second step and isolated by separation over a MACS Column, which is placed in the magnetic field of a MACS Separator. The flow-through contains the enriched MZ B cells. After removing the column from the magnetic field, the magnetically retained FO B cells can be eluted as the positively selected cell fraction.

Columns
LS or autoMACS® Columns.

Overview
The Marginal Zone and Follicular B Cell Isolation Kit has been developed for the isolation of marginal zone (MZ) B cells and follicular (FO) B cells from mouse spleen.