EasySep™ Direct Human Naïve B Cell Isolation Kit

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	B cells were isolated from PBMCs using the EasySep™ Direct Human Naïve B Cell Isolation Kit (STEMCELL Technologies) according to the manufacturer's instructions. B cells were centrifuged (500 × g for 5 minutes), the supernatant was removed, protein was isolated, and western blotting performed as described previously.

Protocol tips
B cells were isolated from PBMCs using the EasySep™ Direct Human Naïve B Cell Isolation Kit (STEMCELL Technologies) according to the manufacturer's instructions. B cells were centrifuged (500 × g for 5 minutes), the supernatant was removed, protein was isolated, and western blotting performed as described previously.

Publication protocol

Total PBMCs were isolated from whole blood from a healthy volunteer and from patients with CLL. Erythrocytes were lysed using RBC Lysis Buffer (Roche, Basel, Switzerland) according to the manufacturer's instructions. Cells were re-suspended in PBS. CLL cells were isolated per the above protocol. B cells were isolated from PBMCs using the EasySep™ Direct Human Naïve B Cell Isolation Kit (STEMCELL Technologies) according to the manufacturer's instructions. B cells were centrifuged (500 × g for 5 minutes), the supernatant was removed, protein was isolated, and western blotting performed as described previously. [18] For patient samples collected in Italy, cells were lysed with RIPA Buffer (Sigma-Aldrich, St. Louis, MO, USA) with a fresh protease and phosphatase inhibitor cocktail (Roche). Western blotting was performed using 30 μg of total protein. Membranes were incubated with 1:250 SR-B1 antibody (Abcam, Cambridge, UK) or 1:50,000 beta actin (Sigma).

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Download the product protocol from STEMCELL technologies for EasySep™ Direct Human Naïve B Cell Isolation Kit below.

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