RosetteSep HLA Myeloid Enrichment Kit

STEMCELL Technologies' negative-selection cell enrichment kit is designed for the in vitro enrichment of myeloid (CD33+) cells from human whole blood, with an intended application in lineage-specific chimerism analysis. The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming immunorosettes whose increased density causes them to pellet along with free RBCs during centrifugation over a density gradient medium. The desired cells remain unlabeled and are collected at the plasma–density medium interface as a highly enriched population.

The companion density medium has a density of 1.085 g/mL and is formulated to optimize myeloid cell recovery. Typical purity of enriched myeloid cells from samples of normal, healthy adults is > 85%. Each lot is manufactured using aseptic technique and sterility tested according to USP methods. The kit is for in vitro diagnostic use by professional users and should be handled by trained personnel observing good laboratory practice.

The antibody cocktail is stored at 2 - 8°C (it may be shipped at 15 - 25°C but must not be frozen) and the density medium at 15 - 25°C. The standard procedure adds cocktail at 50 μL/mL of whole blood, incubates for 20 minutes at room temperature, dilutes the sample, layers it on the density medium, and centrifuges at 330 x g for 25 minutes at room temperature with the brake off. Recommended wash and dilution medium is Dulbecco's PBS with 2% fetal bovine serum, with or without 1 mM EDTA.