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C0773

Sigma-Aldrich

Collagenase from Clostridium histolyticum

high purity, purified by chromatography, Type VII, ≥4 FALGPA units/mg solid, lyophilized powder, ≥700 CDU/mg solid (CDU = collagen digestion units)

Synonym(s):
Clostridiopeptidase A
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.54

Quality Level

type

Type VII

form

lyophilized powder

specific activity

≥4 FALGPA units/mg solid
≥700 CDU/mg solid (CDU = collagen digestion units)

mol wt

68-130 kDa

purified by

chromatography

composition

Protein, ≥85% biuret

solubility

TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C

foreign activity

caseinase and clostripain ≤1 units/mg protein

storage temp.

−20°C

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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.

Packaging

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

Biochem/physiol Actions

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.
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.

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.

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.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificate of Analysis

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Certificate of Origin

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Product Information Sheet

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