RoboSep Mouse CD4+CD62L+Tcell Iso Kit

This kit from STEMCELL Technologies enables the isolation of highly purified naïve CD4+ T cells (CD4+CD62L+) from mouse splenocytes or other single-cell suspensions through a column-free, two-step immunomagnetic procedure. In the first step, naïve CD4+ T cells are pre-enriched using the EasySep™ Mouse Naïve CD4+ T Cell Pre-Enrichment Cocktail, which contains antibodies recognizing specific cell surface markers. Cells pre-labeled with CD62L PE are then selected using the EasySep™ PE Selection Cocktail. These cocktails label cells with antibodies that link to magnetic particles, and separations are performed using an EasySep™ magnet without columns. The manufacturer notes that when using single-cell suspensions from tissue types other than spleen, the kit may require optimization.

Up to 97% purity is achievable. Starting with mouse splenocytes, the naïve CD4+ T cell content (CD4+CD44low CD62Lhigh) of the final isolated fraction typically ranges from 91.7% to 96.7%; in one example, the start and final isolated fractions measured 17.9% and 92.4%, respectively. The kit is supplied for processing 1 x 10^9 cells and supports both the automated RoboSep™ protocol and manual EasySep™ magnet workflows. All kit components should be stored at 2 - 8°C and must not be frozen.

Isolated cells are immediately available for downstream applications such as flow cytometry, cell culture, or DNA/RNA extraction. Purity can be assessed by flow cytometry using the Anti-Mouse CD4 Antibody, Clone RM4-5, together with the Anti-Mouse CD44 Antibody, Clone IM7; the positively selected CD62L+ cells are already PE-labeled and can be assessed directly by flow cytometry.