Synonyms: dissociating enzyme; neutral protease; tissue dissociating
Specific Activity: ≥0.8 units/mg protein (37 °C, casein as substrate, pH 7.5)
UNSPSC Code: 12352200
RIDADR: NONH for all modes of transport
Features and Benefits:
• Rapid, effective, yet gentle agent that liberates cells with minimal cell damage
• Maintains cell membrane integrity
• Non-mammalian source - free of mycoplasma and animal virus contamination
• Extremely stable to influences of temperature, pH, and interference by serum components
• Easily inactivated by chelating agents or by dilution
• Delivers higher activity and convenience
Application:
Dispase® is used for the preparation of cells from a wide variety of different tissues and organs. The enzyme has proven to be a rapid, effective, yet gentle agent for the disruption of extracellular matrix of tissues for releasing single cells for cell culture (primary cell culture) or harvesting cells already in culture to transfer to new substrate (secondary culture). Furthermore, it is used to prevent unwanted clumping of cells cultured in suspension.
General description:
Dispase is a neutral, rapid-acting, gentle protease that separates intact epidermis from dermis. It also separates cultured intact epithelial cells from the substratum. It cleaves the basement membrane zone and leaves the epithelial cells intact.
Other Notes:
For life science research only. Not for use in diagnostic procedures.
Preparation Note:
Activator: Optimal Ca2+ concentration is 2 mM. The enzyme preparations contain enough Ca2+ for optimal activity.
Inhibitors: EDTA, EGTA, Hg2+, other heavy metals. Dispase is not inhibited by serum.
Working concentration: 0.6 to 2.4 U/ml
Working solution: Preparation of stock and working solutions:
To produce a 10 mg/ml stock solution, dissolve the lyophilized Dispase® II enzyme in HEPES-buffered saline (50 mM HEPES/KOH pH 7.4, 150 mM NaCl). To produce the working solution, dilute the above stock solution with the culture medium for the isolated cells, at a final concentration of 0.6 to 2.4 U/ml. Note that concentrations higher than 2.4 U/ml are not recommended. For best results, filter the working solution using a 0.22 μm filter membrane.
Storage conditions (working solution): -15 to -25 °C
The reconstituted stock solution is stable at 2 to 8 °C for 2 weeks. For storage up to 2 months the stock solution should be frozen in aliquots. Avoid repeated freezing and thawing! The working solution diluted with PBS is stable at 2 to 8 °C for 3 days.
Specificity:
Dispase is a nonspecific metalloprotease.
Legal Information:
Dispase is a registered trademark of Godo Shusei Co., Ltd.