RosetteSep Human CD4+ Enrichment, 2 mL

From STEMCELL Technologies, this enrichment cocktail uses an immunodensity procedure for the isolation of untouched CD4+ T cells directly from human whole blood by negative selection. Unwanted cells are crosslinked to red blood cells (RBCs) present in the sample, so that the target CD4+ T cell population is purified during standard density gradient centrifugation using media such as Lymphoprep™, Ficoll-Paque™, or RosetteSep™ DM-L Density Medium.

The workflow involves incubating the whole blood sample with the appropriate RosetteSep™ cocktail for 20 minutes, layering it on a density gradient medium, and centrifuging for 20 minutes. The untouched CD4+ T cells are then collected from the interface between the plasma and the density gradient medium. The procedure can be combined with SepMate™ PBMC isolation tubes to increase sample throughput and reduce handling errors associated with improper sample layering or PBMC harvesting.

According to the manufacturer's FACS analysis starting with fresh human whole blood, the CD4+ (CD3+CD4+) T cell content of the enriched fraction is typically 94.7 ± 2.4% (mean ± SD), with the purities of the start and final enriched fractions reported as 15.1% and 95.0%, respectively. The reagent contains a mouse monoclonal IgG2b antibody against human, rhesus, and cynomolgus CD4, and is designed for use as part of a workflow for immunomagnetic negative selection of untouched human CD4+ T cells and for density gradient centrifugation in in vitro diagnostic (IVD) applications.