EasySep™ Human B Cell Isolation Kit

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	B cells were purified from mouse spleen or human PBMC using magnetic negative selection kits (StemCell Technologies).

Protocol tips
B cells were purified from mouse spleen or human PBMC using magnetic negative selection kits (StemCell Technologies).

Publication protocol

B cells were purified from mouse spleen or human PBMC using magnetic negative selection kits (StemCell Technologies). Cells were plated into 96-well round-bottom dishes at 1 x 105 cells/well with concentrations of mAb as described for individual experiments. In some cases, 1 x 105 irradiated CHO-K1 or 293F cells transfected with human FcγRIIB (White et al., 2011) were also added. To assess activation, cells were photographed (Olympus CKX41 microscope with CC12 soft imaging system) after 48 hour incubation and activation marker expression analysed by flow cytometry (FACSCalibur, BD Biosciences). Proliferation was assessed by [methyl-3H] thymidine (PerkinElmer, Cambridge, UK) incorporation after 5 days of culture, as described (White et al., 2011).

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