The video explains a way to purify B-cells by immunomagnetic targeting and separation from peripheral blood mononuclear cells suspension. The method involve labeling unwanted cell types with specific antibodies or ligands (linked to magnetic particles) targeting specific cell surface proteins, to be depleted from the heterogenous cell suspension by magnetic separation.
Protocol
1. Purification of Leukemic B Cells by Negative Isolation Resuspend 107 PBMCs/ml in PBS 0.1 % bovine serum albumin (BSA) 2 mM EDTA, pH 7.4 (isolation buffer). Add 10 µg of mouse monoclonal anti-CD3, -CD14 and -CD16 primary antibodies for 107 cells and incubate for 30 min at 4 °C. Wash the cells with the isolation buffer and centrifuge at 500 x g for 5 min at 4 °C. Resuspend the cells in the isolation buffer at 107 PBM…