COLLAGÈNE / disquette ROCHE

Non disponible en dehors du Royaume-Uni et de l'Irlande

Application

Collagenase/Dispase^® is used for the preparation of cells from a wide variety of different tissues and organs.

Components

EC 3.4.24.4 & 3.4.24.3

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. 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. 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.

Legal Information

Dispase is a registered trademark of Godo Shusei Co., Ltd.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Collagenase from Vibrio alginolyticus, Dispase^® from Bacillus polymyxa

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).

Specificity

Collagenase degrades native collagen. Dispase^® is a nonspecific protease.

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.