GCB cells from the spleens of SRBC immunized mice were sorted after enrichment of by negative selection, using the MagniSort Mouse B cells Enrichment Kit (ThermoFisher Scientific), with addition of anti-IgD-biotin antibody to deplete IgD+ naïve cells. GC B cell pools were sorted using GL7+FAS+ (Cγ1-cre+/− control mice) and GL7+FAS+ DAF+ (DAF-TMCγ1 mice) in a BD FACS Aria ™ III. DAF expression was used in the GL7+/FAS+ fraction to confirm transgene expression and successful recombination. In other analyses, cells in S phase were in vivo labeled for 2 h upon i.v. injection with 1 mg of 5-ethynyl-20-deoxyuridine (EdU)65 (link). Cells in S phase were gated as BrdU+ 7AADint.
Magnisort mouse b cells enrichment kit
The MagniSort Mouse B cells Enrichment Kit is a magnetic-based cell separation tool designed to enrich for mouse B cells from a heterogeneous cell population. The kit utilizes antibody-coated magnetic beads to specifically target and isolate the B cell population.
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2 protocols using magnisort mouse b cells enrichment kit
Isolation and Analysis of Murine Splenic GCB Cells
Isolation and Analysis of Murine Splenic GCB Cells
GCB cells from the spleens of SRBC immunized mice were sorted after enrichment of by negative selection, using the MagniSort Mouse B cells Enrichment Kit (ThermoFisher Scientific), with addition of anti-IgD-biotin antibody to deplete IgD+ naïve cells. GC B cell pools were sorted using GL7+FAS+ (Cγ1-cre+/− control mice) and GL7+FAS+ DAF+ (DAF-TMCγ1 mice) in a BD FACS Aria ™ III. DAF expression was used in the GL7+/FAS+ fraction to confirm transgene expression and successful recombination. In other analyses, cells in S phase were in vivo labeled for 2 h upon i.v. injection with 1 mg of 5-ethynyl-20-deoxyuridine (EdU)65 (link). Cells in S phase were gated as BrdU+ 7AADint.
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