EasySep Ms Pan-ILC Enrich Kit

STEMCELL Technologies offers a column-free immunomagnetic negative selection kit for enriching untouched group 1, 2, and 3 innate lymphoid cells (ILC1, ILC2, and ILC3) from single-cell suspensions of mouse lung, lymph node, or other tissue samples. The procedure can be completed in as little as 16 minutes, with isolated cells immediately available for downstream applications such as flow cytometry or cell sorting.

The kit targets non-ILCs for removal with biotinylated antibodies recognizing specific cell surface markers. Unwanted cells are labeled with magnetic particles and separated without columns using an EasySep™ magnet, and desired cells are simply poured off into a new tube. The Enrichment Cocktail is a combination of monoclonal antibodies in PBS and 0.1% BSA, while the Streptavidin RapidSpheres™ component is a suspension of magnetic particles in water. Both components should be stored at 2 - 8°C.

From naïve mouse lung single-cell suspensions, the total ILC content (CD45+Lin-CD127+) of the enriched fraction typically ranges from 3.1 - 7.6%, while starting fractions typically range from 0.3 - 0.6%. From naïve mouse lymph node single-cell suspensions, the total ILC content of the enriched fraction typically ranges from 21.1 - 45.2%, with starting fractions typically ranging from 0.3 - 0.4%. ILCs are defined as CD45-positive, lineage-negative, and CD127-positive, with ILC1s characterized as CD278-CD117-, ILC2s as CD278+CD117+/-, and ILC3s as CD278-CD117+. The recommended medium is EasySep™ Buffer, or PBS containing 2% FBS and 1 mM EDTA, free of Ca++, Mg++, and biotin. Use with single-cell suspensions from other tissue types may require optimization.