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C7926 Sigma

Collagenase from Clostridium histolyticum

Sigma Blend Type F, ≥2.0 FALGPA units/mg solid

Synonym: Clostridiopeptidase A

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Description

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 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. The pH optimum is 6.3-8.8. The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA)4; β-mercaptoethanol; glutathione, reduced; thioglycolic acid, sodium; and 2,2′-dipyridyl; 8-hydroxyquinoline are known to inhibit the enzyme activity.

Packaging

1, 5 g in glass bottle

100, 500 mg in glass bottle

25 mg in poly bottle

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.

Application

Collagenase from Clostridium histolyticum, or Clostridiopeptidase A, has been used in a study to assess contact dermatitis with clostridiopeptidase A contained in Noruxol ointment. Clostridiopeptidase A has also been used in a study to investigate the early and successful enzymatıc debridement via collagenase application to pinna in a preterm neonate.

The enzyme has been used in the isolation of mast cells from adult mice. This study generated a large number of connective tissue-type mast cells by the culture of murine fetal skin cells. The product has also been used in the in vitro biodegradation study of Bombyx mori silk fibroin fibers and films, by exposing them to the enzyme for varied time periods.

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Laboratory Gloves

Biomedical Applications
Safety & Documentation

Safety Information

Symbol 
Signal word 
Danger
Hazard statements 
Precautionary statements 
Personal Protective Equipment 
RIDADR 
NONH for all modes of transport
WGK Germany 
1

Documents

Certificate of Analysis

Certificate of Origin

Protocols & Articles

Articles

Collagenase Tissue Digestion/Dissocation Trouble Shooting and References

* The separately prepared collagenase and protease enzymes in the “Sigma Blend” products (Cat. No. C7926, C8051 or C8176) give reproducible control of how much of each is used.
BioFiles 2006, 1.2, 6.
Keywords: Adhesion, Apoptosis, Applications, Buffers, Catalog, Cell biology, Digestions, Methods

Protocols

Enzymatic Assay of Collagenase

The continuous spectrophotometric rate determination (A345, Light path = 1 cm) is based on the following reaction:
Keywords: Extinction coefficient

Related Content

Enzymes & Proteins

Application Index | Enzyme Index | Substrate Index | Inhibitor Index | Cofactor Index | Lectin Index
Keywords: Cell signaling, Diagnostic, Drug discovery, Molecular biology

Peer-Reviewed Papers
15

References

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