RosetteSep HLA Lymphoid Enrichment Kit

The RosetteSep™ HLA Lymphoid Cell Enrichment Kit from STEMCELL Technologies is an in vitro diagnostic kit designed for the negative selection enrichment of lymphoid cells (CD3+) directly from human whole blood, supporting applications such as lineage-specific chimerism analysis. The antibody cocktail crosslinks unwanted cells to multiple red blood cells, forming immunorosettes that pellet along with free red blood cells during density gradient centrifugation. The desired lymphoid cells remain unlabeled and are collected as a highly enriched population at the interface between the plasma and the density gradient medium.

The protocol uses 10 mL of whole blood per test, combined with the cocktail at 50 μL/mL of whole blood and incubated for 20 minutes at room temperature (15 - 25°C). After dilution with an equal volume of recommended medium, the sample is layered over the density gradient medium and centrifuged at 330 x g for 25 minutes at room temperature (15 - 25°C) with the brake off. The density gradient medium supplied has a density of 1.081 g/mL. Reported purity is > 85% for lymphoid cells (CD3+), based on results from normal, healthy adult samples.

This immunodensity cell isolation platform requires no columns or magnets. The cocktail should be stored at 2 - 8°C, and the density medium at 15 - 25°C, protected from direct light. The kit is intended for professional users only and should be handled by trained personnel observing good laboratory practices.