Immunomagnetic selection of human CD4+CD127lowCD25+ regulatory T cells using particle release technology

• Highly purified human CD4+CD127lowCD25+ Tregs isolated in less than 1 hour
• No-wash removal of EasySep™ Releasable RapidSpheres™
• Optional isolation of CD4+CD25- responder T cells from the same sample

This kit isolates human CD4+CD127lowCD25+ regulatory T cells (Tregs) from fresh or previously frozen peripheral blood mononuclear cells (PBMCs) or leukapheresis samples. First, CD25+ cells are isolated by column-free immunomagnetic positive selection using EasySep™ Releasable RapidSpheres™. Then, bound magnetic particles are removed from the EasySep™-isolated CD25+ cells, and unwanted non-Tregs are targeted for depletion. The final isolated fraction contains highly purified CD4+CD127lowCD25+ cells that express high levels of FOXP3 and are immediately ready for downstream applications. An optional protocol allows for the isolation of CD4+CD25- responder T cells in parallel for use in functional studies. Following cell isolation with this EasySep™ kit, antibody complexes remain bound to the cell surface and may interact with Brilliant Violet™ antibody conjugates, polyethylene glycol-modified proteins, or other chemically related ligands.