RosetteSep HLA Total Lymph Cktl, 40mL

A lymphocyte enrichment cocktail from STEMCELL Technologies isolates untouched lymphocytes directly from human whole blood by negative selection. The cocktail uses Tetrameric Antibody Complexes that recognize CD16, CD36, CD66b, and glycophorin A on red blood cells, crosslinking unwanted cells to multiple RBCs to form immunorosettes. During centrifugation over a buoyant density gradient medium with a density of 1.077 g/mL, the immunorosetted cells pellet with the free RBCs, while the desired lymphocytes collect as a highly enriched population at the plasma–density gradient interface.

The protocol is fast and easy-to-use, requiring no special equipment or training. The cocktail is added at 40 μL/mL of whole blood, incubated at room temperature (15–25°C) for 20 minutes, then diluted with an equal volume of recommended medium and centrifuged at 1200 x g for 20 minutes with the brake off. Recommended materials include RosetteSep™ DM-L Density Medium or Lymphoprep™ for the density gradient step, with Dulbecco's PBS containing 2% fetal bovine serum as the wash medium. Enriched cells are immediately suitable for downstream applications such as flow cytometry, cell culture, or DNA/RNA extraction, including flow cytometry crossmatch assays.

The product is a combination of monoclonal antibodies in PBS, stored at 2–8°C and not frozen, and is stable until the expiry date on the label. Each lot is sterility tested according to USP methods and performance tested in cell separation assays using human whole blood. Reported purity for enriched total lymphocytes (T [CD3+] and B [CD19+] cells) is greater than 80%, with CD3+CD16+ NK cells removed during the procedure. The cocktail has been CE Marked in the European Union under the 98/79/EC In Vitro Diagnostic Device Directive. The product is intended for research use only and is not intended for human or animal diagnostic or therapeutic uses unless otherwise stated.