Mouse Neutrophil Negative Selection Kit

STEMCELL Technologies provides an EasySep™ kit that isolates untouched, highly purified neutrophils from mouse bone marrow or peripheral blood by immunomagnetic negative selection. The fast, easy-to-use, and column-free procedure labels unwanted cells (T cells, B cells, NK cells, dendritic cells, progenitors, monocytes, and granulocytes) with biotinylated antibodies and magnetic particles; desired cells are simply poured off into a new tube. Isolated cells are immediately available for downstream applications such as flow cytometry, cell culture, or cell-based assays, with up to 93% (blood) and 91% (bone marrow) purity reported.

The kit contains four components: EasySep™ Mouse Neutrophil Enrichment Cocktail (a combination of monoclonal antibodies in PBS and 0.1% BSA), EasySep™ Biotin Selection Cocktail (a combination of monoclonal antibodies in PBS), EasySep™ D Magnetic Particles (a suspension of magnetic particles in TBS), and EasySep™ Mouse FcR Blocker (a combination of monoclonal antibodies in PBS, 0.1% BSA, and less than 0.1% sodium azide). All components are stored at 2 - 8°C and are stable until the expiry date on the label; components may be shipped at room temperature (15 - 25°C). The kit is compatible with the EasySep™ Magnet (0.5 - 2 mL samples, 3 minutes in magnet), "The Big Easy" EasySep™ Magnet (0.5 - 6.5 mL samples, 5 minutes in magnet), and the fully automated RoboSep™ system (0.5 - 6.5 mL samples), processing up to 1 x 10^9 cells.

Sample preparation for bone marrow involves flushing cells from femur and tibia using a 23 gauge needle and straining through a 70 μm mesh; peripheral blood is lysed with Ammonium Chloride Solution (1 part blood to 9 parts solution, 15 minutes on ice). Recommended medium is EasySep™ Buffer, RoboSep™ Buffer, or PBS with 2% FBS and 1 mM EDTA (Ca++ and Mg++ free). Purity is assessed by flow cytometry for CD11b+Ly-6G+ cells using the M1/70 and 1A8 antibody clones. The CD11b+Ly-6G+ cell content of the enriched fraction is typically 88.2 ± 3.2% for bone marrow and 88.6 ± 4.9% for blood (mean ± SD using the purple EasySep™ magnet). For research use only; not intended for human or animal diagnostic or therapeutic uses.