A cell detachment solution of proteolytic and collagenolytic enzymes. Useful for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware, including SmartPlastic. ACCUTASE does not contain mammalian or bacterial derived products. Because the reagent does not contain universally proteolytic enzymes it may not release all cell/tissue types uniformly. Some optimization of cell type and passage number, as well as, incubation time and reagent strength typically needs to be performed to optimize the reagents effectiveness. ACCUTASE has been shown to be effective on a wide variety of cell types including: fibroblasts, keratinocytes, vascular endothelial cells, hepatocytes, vascular smooth muscle cells, hepatocyte progenitors, primary chick embryo neuronal cells, bone marrow stem cells, adherent CHO and BHK cells, macrophages, 293 cells, L929 cells, immortalized mouse testicular germ cells, 3T3, Vero, COS, HeLa, NT2, MG63, M24 and A375 metastatic melanoma, gliomas U251 and D54, HT1080 fibrosarcoma cells, and Sf9 insect cells. Each lot of ACCUTASE is tested for Sterility (by USP membrane filtration method), enzymatic activity (tested with synthetic chromagenic tetrapeptides) and cell detachment from tissue culture plastic.
Application: Cell Detachment 1. Thaw ACCUTASE® at room temperature. 2. Wash plate, flask or beads with sterile PBS. 3. Add ACCUTASE to culture dish or flask using aseptic procedures at 10 ml per 75-cm2 surface area. 4. Return culture to 37C incubator and allow cells to detach (5-10 minutes). 5. Count cells and passage as usual. No additional washes or enzyme inhibitors are required Optimal incubation times and reagent strength needs to be determined for each cell/tissue type tested by the enduser.
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