RosetteSep Multi Myeloma Enrch Cocktail

A negative-selection enrichment cocktail from STEMCELL Technologies designed to isolate untouched human multiple myeloma cells (B cells and plasma cells) from fresh human whole bone marrow. The cocktail crosslinks unwanted cells to red blood cells (RBCs) to form immunorosettes, which pellet with free RBCs during density gradient centrifugation; the desired cells remain unlabeled and are collected at the interface between the plasma and the density gradient medium. No special equipment or training is required, and isolated cells are immediately available for downstream applications such as flow cytometry, cell culture, and DNA/RNA extraction.

The reagent is supplied as a combination of monoclonal antibodies in phosphate-buffered saline (PBS). It is recommended for use only with samples containing malignant plasma cells, and an antibody against CD56 is not included in the cocktail. Bone marrow samples should not exceed a nucleated cell concentration of 5 × 10^7 cells/mL, and when enriching from alternative sources such as buffy coat, RBCs must be present at a ratio of at least 100 RBCs per nucleated cell. The recommended medium is Dulbecco's Phosphate Buffered Saline with 2% fetal bovine serum, and the density gradient medium should have a density of 1.077 g/mL. Typical purity is reported as > 95% when measured by anti-CD138 labeling.

The workflow uses 50 µL of cocktail per mL of sample with a 20-minute room-temperature (15 – 25 °C) incubation, followed by layering over density gradient medium and centrifugation at 1200 × g for 20 minutes with the brake off. Enriched cells are washed at 300 × g for 10 minutes and can be assessed for purity using an anti-human CD138 antibody (Syndecan-1), Clone MI15. Components may be shipped at room temperature (15 – 25 °C) but should be stored at 2 – 8 °C and not frozen; a precipitate may be observed in the vial that does not affect performance. The product is for research use only and is not intended for human or animal diagnostic or therapeutic uses.