RosetteSep Debulking for Freezing, 2mL

From STEMCELL Technologies, this antibody cocktail enriches hematopoietic stem and progenitor cells directly from human whole cord blood by negative selection. Lineage-positive cells are targeted for removal using monoclonal antibodies recognizing CD2, CD14, CD19, CD66b, and glycophorin A on red blood cells. The antibody cocktail crosslinks unwanted cells to multiple red blood cells, forming immunorosettes. This increases the density of the unwanted (rosetted) cells, such that they pellet along with the free RBCs when centrifuged over a density gradient medium. The desired cells remain unlabeled and are collected as a highly enriched, untouched, and viable population at the interface between the plasma and the density gradient medium.

Designed to be fast and easy to use, the cocktail requires no special equipment or training and can be combined with SepMate™ for consistent, high-throughput sample processing. Isolated cells are immediately available for downstream applications, such as flow cytometry, cell culture, or DNA/RNA extraction.

For optimal performance, whole cord blood collected within the last 24 hours and stored at room temperature (15–25°C) is recommended, with nucleated cell concentrations not exceeding 5 x 10^7 cells/mL. The cocktail is added at 50 µL per mL of sample, with incubation at room temperature for 20 minutes in a standard tube protocol or 10 minutes when using SepMate. Centrifugation is performed at 1200 x g for 20 minutes with brake off, or 1200 x g for 10 minutes with brake on for SepMate. The recommended density gradient medium is Lymphoprep™ (density 1.077 g/mL) and the recommended medium is Dulbecco's Phosphate Buffered Saline with 2% Fetal Bovine Serum. Starting with fresh human cord blood, the CD34+ content of the de-bulked fraction is typically 5 ± 1%. The cocktail is supplied as a combination of monoclonal antibodies in PBS and should be stored at 2–8°C without freezing; it is stable until the expiry date on the label.