EasySep Direct Hu Total Lymph-RoboSep

From STEMCELL Technologies, this column-free immunomagnetic negative selection kit is designed for use with the RoboSep™ automated platform, enabling isolation of highly purified lymphocytes directly from human whole blood, buffy coat, or spleen. The workflow targets non-lymphocytes for removal using a cocktail of monoclonal antibodies together with EasySep™ Direct RapidSpheres™, with unwanted cells separated on an EasySep™ magnet and the desired cells simply collected untouched into a new tube.

The kit is formulated for processing 100 mL of whole blood and offers >99.9% red blood cell depletion without the need for density gradient centrifugation, sedimentation, or lysis, with up to 96% purity of isolated cells. In a typical isolation profile from normal healthy donor whole blood, the total lymphocyte (CD3-CD19+ and CD3+) content of the non-lysed final isolated fraction is 96.7 ± 1.5% when gated on CD45, or 95.8 ± 2.2% when not gated on CD45. Isolated cells are immediately suitable for downstream applications such as flow cytometry, culture, or DNA/RNA extraction. The kit is also highlighted for use in flow cytometric crossmatch (FCXM) assays, where highly enriched lymphocyte preparations have been shown to improve donor-specific antibody (DSA) detection compared with whole leukocyte cell preparations.

Sample preparation guidance recommends heparin or acid citrate dextrose (ACD) as anticoagulants, with D-PBS without calcium and magnesium as the recommended medium. Recovery of the desired isolated cells is reported to decrease with samples that are older than 24 hours. Kit components should be refrigerated upon receipt and stored at 2 - 8°C.