RoboSep Mouse CD25 Treg Positive Sel Kit

From STEMCELL Technologies, this immunomagnetic positive selection kit enables researchers to isolate highly purified CD25+ regulatory T cells from mouse splenocytes or other single-cell suspensions. The kit targets CD25+ cells using a PE-labeled antibody recognizing the CD25 surface marker; desired cells are labeled with antibodies and magnetic particles and separated without the need for columns using an EasySep™ magnet, so unwanted cells can simply be poured off while the desired cells remain in the tube.

Isolated cells are immediately available for downstream applications such as flow cytometry, culture, or DNA/RNA extraction. The kit is supplied with an EasySep™ Mouse CD25 Regulatory T Cell Positive Selection Cocktail, an EasySep™ PE Selection Cocktail, EasySep™ Dextran RapidSpheres™, and an EasySep™ Mouse FcR Blocker, and is rated for processing 1 x 10^9 cells. Protocols are provided for manual use with the EasySep™, "The Big Easy", and EasyEights™ magnets, as well as for fully automated separation on the RoboSep™ instrument.

Starting with mouse splenocytes, the CD25+ regulatory T cell content (CD4+CD25+FOXP3+) of the isolated fraction typically ranges from 80 - 93%, with example purities of 2.1% in the start fraction and 87.5% in the final isolated fraction. For purity assessment by flow cytometry, STEMCELL Technologies recommends an Anti-Mouse CD4 Antibody, Clone RM4-5, alongside an anti-mouse FOXP3 antibody; note that the CD25+ cells have already been PE-labeled during the selection procedure.