D4693
Dispase® II
protease
Synonym(s):
Protease from Bacillus polymyxa
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About This Item
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Quality Level
form
lyophilized solid
specific activity
≥0.5 units/mg solid
technique(s)
cell culture | mammalian: suitable
single cell analysis: suitable
solubility
10 mM NaAc (pH 7.5) and 5 mM CaAc: soluble
storage temp.
2-8°C
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Application
The enzyme from Sigma has been used in developing a protocol for ex vivo culture of mouse embryonic mammary buds. It has been used in the treatment of rat heart pieces during the isolation of mitochondria from rat heart. It has also been used for the isolation of dental pulp stem cells (DPSCs) by enzymatic hydrolysis. These cells have been compared with DPSCs isolated by explant method to analyse their stem cell and differentiation properties.
Dispase® II has been used for Fluorescence-Activated Cell Sorting (FACS). It has also been used for separating visceral yolk sac layers.
Suitable for use in preparation of single cell suspension for sequencing.
Dispase® II has been used for Fluorescence-Activated Cell Sorting (FACS). It has also been used for separating visceral yolk sac layers.
Suitable for use in preparation of single cell suspension for sequencing.
Biochem/physiol Actions
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. This protease cleaves fibronectin and type IV collagen, but not laminin, type V collagen, serum albumin, or transferrin. Dispase® II is specific for the cleavage of Leucine-Phenylalanine bonds. Ca2+, Mg2+, Mn2+, Fe2+, Fe3+ and Al3+ activate the enzyme. EDTA, EGTA, Hg2+ and other heavy metals inhibit the enzyme activity. The enzyme contains 1g-atom of zinc per g-mol of purified enzyme. If this zinc component is removed by chelating agents such as EDTA or EGTA, an inactive apoenzyme is obtained. The enzyme is not inhibited by serum.
Unit Definition
One unit will hydrolyze casein to produce color equivalent to 1.0 μmole (181 μg) of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent), unless otherwise indicated.
Physical form
lyophilized powder containing calcium acetate and milk sugar
Preparation Note
A 50 U/mL stock solution may be prepared by dissolving the powder in a buffer containing 10 mM NaAc (pH 7.5) and 5 mM CaAc. It should be stored at 4 °C.
Analysis Note
The enzyme can be initially dissolved in 50 mM Hepes/KOH pH 7.4, 150 mM NaCl at 10 mg/ml. Further dilutions should be made in the cell culture media most appropriate for the cells being dissociated. Dispase is a metalloproteinase and is activated by divalent cations including Ca2+ , Mg2+, Mn2+ and Fe2+.
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 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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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