Akadeum’s Mouse B Cell Isolation Kit delivers a high-purity population of mouse B cells that are ready for downstream use. Use of these microbubbles to enrich mouse splenocyte samples for B cells prior to flow sorting can greatly reduce final sort times, helping to isolate more cells for better science. Akadeum’s microbubble workflow is exceptionally gentle on cells, helping to maintain the health and physiology of immune cells during enrichment.
Mechanism of Action: Akadeum’s Mouse B Cell Isolation Kit uses a negative selection method to remove unwanted cell populations from the sample, effectively enriching for the B cells of interest while maintaining cellular physiology. The undesired (non-B) cells are labeled with Akadeum’s optimized cocktail of biotinylated antibodies targeting CD3, CD4, CD8a, CD11b, CD11c, CD49b, Gr-1 and TER-119. Streptavidin-coated microbubbles are then mixed into the sample, binding to the labeled cells and floating them to the top of the sample container for removal.
How it Works: First, Akadeum’s biotinylated antibody cocktail is added to the sample to label the contaminating cells. Next, the microbubbles are mixed into the sample where the seek out and bind to the unwanted cells. Once bound, they quickly and gently float the undesired cells to the surface. An optional centrifugation step pellets the B cells, and the microbubbles, microbubble-bound cells, and supernatant are easily aspirated away leaving the untouched and enriched B cells happy, healthy, and ready for downstream use.
- Storage: Store microbubbles, antibody cocktail, and Separation Buffer at 4°C
- Cell Type: Naïve / resting B cells
- Sample Species and Source: Mouse spleen. Lysis of red blood cells prior to use is strongly encouraged.
- Cell Separation Method: Negative Selection
- Capacity: 1 billion cells
This kit comes with:
- Akadeum’s BACS™ Streptavidin Microbubbles
- Akadeum’s Mouse B Cell Antibody Cocktail
- Separation Buffer (Ca2+ and Mg2+ free PBS containing 2 mM EDTA, 0.5% biotin-free BSA, and 0.09% sodium azide)
- 5mL Eppendorf Tubes
For best results, the following supplies are recommended:
- Low retention 1mL pipet tips
- Centrifuge with swinging bucket rotor
- Vacuum aspirator