C0773-1.5KU Display Image

Collagenase from Clostridium histolyticum, high purity, purified by chromatography, Type VII, 4-15 FALGPA units/mg solid, lyophilized powder, 1,000-3,000 CDU/mg solid (CDU = collagen digestion units)

Code: C0773-1.5KU D2-231

Application

Collagenase has been used in the preparation of arterial tissue for the study of advanced glycosylation end products (AGE). The enzyme has also been used along wi...


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Application

Collagenase has been used in the preparation of arterial tissue for the study of advanced glycosylation end products (AGE). The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues. It is also effective in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes.

Biochem/physiol Actions

Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(beta-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, beta-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline.

Effective release of cells from tissue requires the action of both collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca2+) per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. Optimal pH ranges from 6.3-8.8. The enzyme recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid.

Packaging

1500, 3000, 7500, 15000, 30000, 60000 units in glass bottle

Physical form

Lyophilized powder containing calcium chloride

Preparation Note

A stock solution may be prepared by dissolving 0.05 - 0.1 mg of collagenase in one mL of 50 mM TES buffer containing 0.36 mM calcium chloride (TESCA), pH 7.4 at 37 °C.

Unit Definition

One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0 μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions. One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of furylacryloyl-Leu-Gly-Pro-Ala per min at 25°C. One Neutral Protease unit hydrolyzes casein to produce color equivalent to 1.0 μmole of tyrosine per 5 hr at pH 7.5 at 37°C. One Clostripain Unit hydrolyzes 1.0 μmole of BAEE per min at pH 7.6 at 25°C in the presence of DTT.

compositionProtein, ≥85% biuret
foreign activitycaseinase and clostripain ≤1 units/mg protein
formlyophilized powder
mol wt68-130 kDa
purified bychromatography
Quality Level200
solubilityTESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C
specific activity≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)
storage temp.−20°C
typeType VII
Cas Number9001-12-1
This product has met the following criteria:



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