RosetteSep Human CD4 Depletion, 10mL

This antibody cocktail from STEMCELL Technologies is designed to deplete CD4+ cells directly from human whole blood. The cocktail crosslinks unwanted cells to multiple red blood cells (RBCs) to form immunorosettes, increasing their density so they pellet along with free RBCs during centrifugation over a density gradient medium. Desired cells are never labeled 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.

Key features include fast and easy-to-use processing with no special equipment or training required, and the recovery of untouched, viable cells. The product can be combined with SepMate™ for consistent, high-throughput sample processing. It is composed of a combination of monoclonal antibodies in PBS and should be stored at 2 - 8°C without freezing. Precipitate may be observed in the cocktail vial but will not affect performance.

For optimal performance, whole peripheral blood collected within the last 24 hours and stored at room temperature (15 - 25°C) should be used. The concentration of nucleated cells in the sample should not exceed 5 x 10^7 cells/mL. The standard protocol uses 50 µL/mL of cocktail with a 20-minute room temperature incubation, followed by centrifugation at 1200 x g for 20 minutes with the brake off. When using SepMate™ tubes, the incubation is reduced to 10 minutes and centrifugation to 1200 x g for 10 minutes with the brake on. Recommended medium is Dulbecco's Phosphate Buffered Saline with 2% Fetal Bovine Serum, and the recommended density gradient medium is Lymphoprep™ or equivalent at 1.077 g/mL. In a representative dataset, the CD4+ content of the start and depleted fractions was 12% and 1.6%, respectively, with a 2.0 log depletion of CD4+ cells. The product is manufactured under an ISO 13485-certified quality management system and is for research use only.