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Dispase II

Cat. No. M21672
Dispase II Structure
Size Price Availability Quantity
50mg USD 60  USD60 In stock
100mg USD 90  USD90 In stock
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Quality Control & Documentation
Biological Activity

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)


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

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.

Chemical Information
CAS Number 42613-33-2
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
Conversion of different model animals based on BSA (PMID: 27057123)
Species Mouse Rat Rabbit Guinea pig Hamster Dog
Weight (kg) 0.02 0.15 1.8 0.4 0.08 10
Body Surface Area (m2) 0.007 0.025 0.15 0.05 0.02 0.5
Km factor 3 6 12 8 5 20
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.

References

[1] Beln Calvo, et al. IBRO Rep. Dissociation of neonatal and adult mice brain for simultaneous analysis of microglia, astrocytes and infiltrating lymphocytes by flow cytometry

[2] Jae-Kyung Lee, et al. Methods Mol Biol. Microglia isolation from adult mouse brain

[3] Mark Tomishima. Splitting hPSCs with Dispase

[4] Laurence S Lim, et al. Mol Vis. Effect of dispase denudation on amniotic membrane

[5] S J Spurr, et al. Invest Ophthalmol Vis Sci. Isolation of corneal epithelium with Dispase II or EDTA. Effects on the basement membrane zone

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