EasySep Hu PE Positive Selection Kit II

STEMCELL Technologies' EasySep™ Human PE Positive Selection Kit II enables isolation of highly purified cells labeled with PE (phycoerythrin)-conjugated antibodies from fresh or previously frozen human peripheral blood mononuclear cells (PBMCs) or other single-cell suspensions by immunomagnetic positive selection. The kit targets cells labeled with PE-conjugated antibodies (not provided), with desired cells tagged by antibodies and magnetic particles and separated without columns using an EasySep™ magnet. Unwanted cells are simply poured off while desired cells remain in the tube, and isolated cells are immediately available for downstream applications such as flow cytometry, cell culture, or cell-based assays.

Components include the EasySep™ PE Selection Cocktail (a combination of monoclonal antibodies in PBS), Anti-Human CD32 (Fc gamma RII) Blocker for Positive Selection, EasySep™ Dextran RapidSpheres™ 50100 (a suspension of magnetic particles in water), and a RoboSep™ Vial for Primary Conjugated Antibody. All components should be stored at 2 - 8°C and not frozen; they may be shipped at room temperature (15 - 25°C). Sample preparation involves preparing a PBMC suspension by centrifugation over a density gradient medium; previously frozen PBMCs should be incubated with DNase I Solution at 100 µg/mL at room temperature for at least 15 minutes prior to labeling. After preparation, cells are resuspended at 1 x 10^8 cells/mL in recommended medium (EasySep™ Buffer, RoboSep™ Buffer, or PBS containing 2% FBS and 1 mM EDTA, free of Ca++ and Mg++).

Among the kit's applications is the enrichment of T cells labeled with PE-conjugated HLA class I-peptide tetramers, or PE-conjugated antibodies that bind to CD134, CD137, or CD154. In internal testing using tetramer-labeled PBMCs, the starting purity of 0.5% rose to 85.4% in the final isolated cells. The protocol is compatible with EasySep™ ("The Big Easy"), EasyEights™, and RoboSep™ magnets, with separation times of 5 or 10 minutes per round depending on the magnet used.