Components

EC 3.4.24.4 & 3.4.24.3

Other Notes

Collagenase from Vibrio alginolyticus, Dispase^® from Bacillus polymyxa

仅用于生命科学研究。不可用于诊断。

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, EGTA
Collagenase/Dispase is not inhibited by serum.

Storage conditions (working solution): -15 to -25°C
The 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 mol tyrosine.

Application

Collagenase/Dispase^® is used for the preparation of cells from a wide variety of different tissues and organs.^[4][5][6]

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]

性质