Description
Accutase™ is a cell detachment solution comprised of collagenolytic and proteolytic enzymes and does not contain mammalian or bacterial derived products. Accutase™ is a replacement for trypsin solution and can be used for the passaging of cell lines. Aditionally, Accutase™ performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Accutase™ has been demonstrated effective in detaching multiple cell types including: human and mouse embryonic and induced pluripotent stem cells, human neural stem cells, fibroblasts, keratinocytes, vascular endothelial cells, hepatocytes, neural cell types, bone marrow derived 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.
Preparation and Storage
The product should be kept undiluted at -20°C for long term storage, and it may be kept undiluted at 4°C for short term storage.
This product is shipped frozen, but may thaw during transit. On receipt, if it remains cool to the touch, it may be stored at 4 °C for up to 2 months or re-frozen and stored at -20 °C for up 2 years.
Product Notices
- Accutase is a registered trademark of Innovative Cell Technologies, Inc.
- mTESR™1 is a trademark of StemCell Technologies.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
To obtain a single-cell suspension: (Use aseptic techniques if you plan on continuing culture of cells.)
1. Wash the cells with PBS at room temperature.
2. Add Accutase™ cell detachment solution to adequately cover the entire area of the culture vessel.
3. Incubate at room temperature for 5 to 10 minutes, or until cells are detached. Cells can also be incubated at 37°C, however enzyme activity will decrease over time.
a. Additional time and/or Accutase™ may be needed to disassociate three-dimensional structures.
4. Triturate the cells to aid in obtaining a single-cell suspension.
a. Additional cell culture medium may be added to assist in obtaining a single-cell suspension.
b. The additional cell culture medium will neutralize the Accutase™ and allow for passaging of cells without additional washes.
5. Remove a small subset of the cell suspension and examine under a microscope to confirm the presence of single cells.
