Collagenase/Dispase®

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] Collagenase is a protease which cleaves the triple-helical protein called collagen. There are three types of tissue collagenases, and these belong to the matrix metalloproteinases (MMP) family.[2] Collagenase obtained from Clostridium histolyticum has a very strong activity, as it digests collagen from both ends, at temperatures as low as 4-10 °C.[3]
• Specificity: Collagenase degrades native collagen. Dispase® is a nonspecific protease.
• Application: Collagenase/Dispase® is used for the preparation of cells from a wide variety of different tissues and organs.[4][5][6]
• Components: EC 3.4.24.4 & 3.4.24.3
• Unit Definition: One unit is the enzyme activity, which liberates under assay conditions within 1 minute Folin-positive amino acids and peptides corresponding to 1 μmol tyrosine.
• Preparation Note: Activator: Ca2+Working concentration: Approximately 1mg/mL (Collagenase: 0.1U/mL, Dispase: 0.8U/mL)Enzyme activity:Collagenase: 0.1 Wünsch U/mg lyophilizate (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)Dispase®: 0.8 Wünsch U/mg lyophilizate (+37°C, casein as substrate, pH 7.5)Contaminating proteolytic activities: <0.1U/mg lyophilizate (hemoglobin as the substrate)Working solution: Preparation of working solution Dissolve enzyme in water to a final concentration of 100mg/mL. Dilute with PBS (Ca2+/Mg2+-free) to a final concentration of 1mg/mL and filter through a sterile 0.2μm membrane filter.Enzyme concentration of the sterile solution: Collagenase, 0.1U/mL PBS; Dispase®, 0.8U/mL PBS.Inhibitors:Collagenase inhibitors: EDTA, EGTACollagenase/Dispase is not inhibited by serum.Storage conditions (working solution): -15 to -25°CThe reconstituted solution is stable at -15 to -25°C.Note: Store in appropriate aliquots and avoid repeated freezing and thawing!
• Reconstitution: Reconstitution in water (100mg/mL). Further dilution with PBS (phosphate buffered saline) (Ca2+/Mg2+-free).
• Storage and Stability: Store dry
• Other Notes: Collagenase from Vibrio alginolyticus, Dispase® from Bacillus polymyxa
• Legal Information: Dispase is a registered trademark of Godo Shusei Co., Ltd.