RosetteSep Hu CD4+CD127lo Cocktail, 10mL

This negative selection enrichment cocktail from STEMCELL Technologies is designed to process up to 200 mL of human whole blood and to isolate untouched CD4+CD127low T cells directly from the sample. The antibody cocktail crosslinks unwanted cells to multiple red blood cells, forming immunorosettes that pellet along with free RBCs during density gradient centrifugation. Desired cells are never labelled with antibody and are collected as a highly enriched population at the interface between the plasma and the density gradient medium, making them immediately available for downstream applications such as flow cytometry, cell culture, or DNA/RNA extraction.

The kit requires no special equipment or training and can be combined with SepMate™ tubes for consistent, high-throughput sample processing. The formulation is a combination of monoclonal antibodies in PBS and should be stored at 2 – 8°C; freezing should be avoided. The cocktail remains stable until the expiry date printed on the label.

For optimal results, whole peripheral blood should be used within 24 hours of collection and kept at room temperature (15 – 25°C). The concentration of nucleated cells in the sample should not exceed 5 x 10^7 cells/mL. The recommended medium is PBS containing 2% fetal bovine serum and 1 mM EDTA, free of Ca²⁺ and Mg²⁺. Density gradient medium should have a density of 1.077 g/mL. The standard protocol uses 50 µL of cocktail per mL of sample, followed by a 20-minute room-temperature incubation. Centrifugation conditions are 400 x g for 30 minutes (brake off) with a standard tube, or 1200 x g for 10 minutes (brake on) when using SepMate™.