MACSprep™ HLA B Cell Isolation Kit, human

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	Following manufacturer's instructions

Protocol tips
Following manufacturer's instructions

Publication protocol

"Drug Minimal Cytotoxic Concentration Assessment
We assessed in vitro minimal anti-CD20 monoclonal antibody cytotoxic concentration. Anti-CD20 monoclonal antibodies (rituximab, obinutuzumab, ocrelizumab and ofatumumab) at 6.25, 12.5, 25, and 50 ng/ml were incubated with 1.5 × 103 purified B-cells (MACSprepTM HLA B Cell Isolation Kit, Milteny Biotec) for 30 min at room temperature in 60-well Terasaki plates (Dutcher© Strasbourg, France) in duplicates of 1 μl per well. Then, 5 μl per well of standard rabbit complement (Cerdarlane© Ontario, Canada) were added for 45 min at room temperature. Dead cells were then revealed after adding 2.5 μl per well of Fluoroquench AO/EB staining/quench (Ingen© Chilly-Mazarine, France) for 10 min in darkness.

Two blinded independent evaluators read the percentage of dead cells using a fluorescent microscope (Axiovert 100 Carl Zeiss© Göttigen, Germany)."

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Manufacturer protocol

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