RosetteSep HLA Gran Dpln Cktl, 40mL

An immunodensity cell separation reagent from STEMCELL Technologies, the RosetteSep™ HLA Granulocyte Depletion Cocktail is designed to deplete granulocytes from human whole blood. The antibody cocktail crosslinks unwanted cells to multiple red blood cells (RBCs), forming immunorosettes. These increase the density of unwanted cells so that they pellet along with free RBCs when centrifuged over a density gradient medium, while the desired mononuclear cells are never labelled with antibody and are collected as a highly enriched population at the plasma : density gradient interface.

The cocktail contains a combination of monoclonal antibodies in phosphate-buffered saline (PBS). It is intended for in vitro diagnostic use by trained professional users. The workflow requires a density gradient medium with a density of 1.077 g/mL, such as Lymphoprep™, and Dulbecco's PBS with 2% fetal bovine serum as the recommended medium. The product is stored at 2 - 8°C and should not be frozen. It is a potential irritant to the eyes, respiratory system, and skin, and may be harmful if ingested; once added to human cells, the suspension should be treated as potentially biohazardous and handled observing good laboratory practice.

According to the manufacturer's directions, the cocktail is added at 40 μL/mL of whole blood, incubated for 20 minutes at room temperature (15 - 25°C), diluted with an equal volume of recommended medium, layered over the density gradient medium, and centrifuged at 1200 x g for 20 minutes at room temperature with the brake off. The enriched cells are then washed and used as desired. Typical purity of the enriched mononuclear cell population is reported as > 90%, although results from individual patient samples may vary. Granulocyte depletion can be monitored by flow cytometry, assessing depletion of high side scatter cells.