Collagenase from Clostridium histolyticum, high purity, purified by chromatography, Type VII, 4-15 F

Collagenase from Clostridium histolyticum, high purity, purified by chromatography, Type VII, 4-15 F

Stock Code: 3579106
Manufacturer Part No: C0773-7.5KU
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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.

Quality Level200
ManufacturerSIGMA-ALDRICH
CompositionProtein, ≥85% biuret
SolubilityTESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C
Formlyophilized powder

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