EasySep Direct Hu PBMC Isolation-RoboSep

This immunomagnetic negative selection kit from STEMCELL Technologies is designed for processing 100 mL leukapheresis samples and isolates highly purified peripheral blood mononuclear cells (PBMCs) directly from leukapheresis, while also being applicable to whole blood, buffy coat, bone marrow, and cord blood. The kit targets granulocytes, platelets, and red blood cells (RBCs) for removal using antibodies that recognise specific cell surface markers; unwanted cells are labelled with antibodies and magnetic particles and separated using an EasySep™ magnet. The column-free workflow achieves 99.9% red blood cell depletion without the need for density gradient centrifugation, sedimentation, or lysis, and the isolated cells remain untouched and immediately available for downstream applications such as flow cytometry, culture, or DNA/RNA extraction.

This format is configured for use on the fully automated RoboSep™ instrument. Sample preparation calls for adding EDTA to the leukapheresis sample to a final concentration of 6 mM before labelling and separation. Recommended media are EasySep™ Buffer, RoboSep™ Buffer, D-PBS (without Ca++ and Mg++), or PBS containing 2% fetal bovine serum and 1 mM EDTA. The automated workflow completes cell isolation in approximately 30 minutes with about 6 minutes of hands-on time, compared with around 20 minutes of cell isolation and about 21 minutes of hands-on time for the manual EasySep™ Direct protocol.

Kit components include an Isolation Cocktail — a combination of monoclonal antibodies in PBS, including an Fc receptor blocking antibody — and RapidSpheres™, a suspension of magnetic particles and monoclonal antibodies in PBS. Components should be stored at 2 - 8 °C, are stable until the expiry date on the label, and may be shipped at room temperature (15 - 25 °C). Compared with density gradient centrifugation, the kit has been shown to deliver equivalent numbers of total mononuclear cells while reducing residual platelets, red blood cells, and granulocytes, and PBMCs isolated with the kit have been demonstrated to proliferate normally in culture and to maintain high RNA integrity.