Dispase II is a neutral protease that hydrolyzes the N-terminal peptide bonds of non-polar amino acid residues. It may be used for separating many tissues and cells grown in vitro. The enzyme is very gentle and does not damage cell membranes. It can also be used to prevent clumping in suspension cultures.
Enzyme activity: DispaseII ≥0.8 U/mg (37°C, using casein as substrate, pH 7.5)
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
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): -20 °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.
|Solubility (25°C)||50 mM HEPES/KOH pH 7.4 150 mM NaCl|
|Storage||Stable for 6-12 months after receipt, store at 2-8°C, dry, sealed|
|Body Surface Area (m2)||0.007||0.025||0.15||0.05||0.02||0.5|
|Animal A (mg/kg) = Animal B (mg/kg) multiplied by||Animal B Km|
|Animal A Km|
For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.
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