Dispase® I (neutral protease, grade I)

Description

• 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.[1]
• Specificity: Dispase® is a non-specific metalloprotease.
• Application: Dispase® is used for the preparation of cells from a wide variety of different tissues and organs.[2][3][4] 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.
• Features and Benefits: Rapid, effective, yet gentle agent that liberates intact cellsMaintains cell membrane integrityNon-mammalian source - free of mycoplasma and animal virus contaminationExtremely stable to influences of temperature, pH, and interference by serum componentsEasily inactivated by chelating agents or by dilutionDelivers higher activity and convenienceContents10 vials with approximately 2mg each (= 20U/vial).
• Specifications: Enzyme activity: =10U/mg (+37°C, casein as substrate, pH 7.5)Inhibitors: EDTA, EGTA, Hg2+, other heavy metals. Dispase is not inhibited by serum.
• Unit Definition: One unit is the enzyme activity which liberates Folin-positive amino acids and peptides from casein corresponding to 1 μmol (181 μg) tyrosine in 1 minute under assay conditions (pH 7.5, +37 °C).A practical comparison of RAS units of Dispase® with those cited in the Japanese literature (where concentrations of 1,000 to 2,000 Dispase® units /ml are not uncommon) suggests one Roche Applied Science unit of Dispase® I equals approximately 416 Japanese units of Dispase®.One unit of Roche Applied Science Dispase® equals 181 protease units (PU) measured as release of amino acids equivalent to 1 μg tyrosine per min and ml at pH 7.5 and +37 °C.
• Physical form: white lyophilizate, before lyophilization filtered through 0.2 μm pore size membrane
• Preparation Note: Activator: Optimal Ca2+ concentration is 2 mM. The enzyme preparations contain enough Ca2+ for optimal activity.Working concentration: 0.6 to 2.4 U/mlWorking solution: Preparation of stock and working solutions:To produce a 10 mg/ml stock solution, dissolve the lyophilized Dispase® I 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 °CThe 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.
• Storage and Stability: Store at 2 to 8 °C. (Store dry!)
• Other Notes: For life science research only. Not for use in diagnostic procedures.
• Legal Information: Dispase is a registered trademark of Godo Shusei Co., Ltd.