04942086001
Roche
Dispase® I (neutral protease, grade I)
lyophilized, ≥10 units/mg protein (20 U/vial (37 °C, casein as substrate, pH 7.5)), optimum pH 6.0-8.5
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About This Item
Recommended Products
biological source
Bacillus polymyxa
Quality Level
form
lyophilized
specific activity
≥10 units/mg protein (20 U/vial (37 °C, casein as substrate, pH 7.5))
packaging
pkg of 10 × ~2 mg
manufacturer/tradename
Roche
concentration
0.6-2.4 U/mL
technique(s)
tissue processing: suitable
color
white
optimum pH
6.0-8.5
solubility
water: soluble
application(s)
life science and biopharma
sample preparation
shipped in
wet ice
storage temp.
2-8°C
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.
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. 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 cells
- 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
Contents
10 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.
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.
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/ml
Working 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 °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.
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® 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 °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.
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.
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
Target Organs
Respiratory system
Storage Class Code
11 - Combustible Solids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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