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C2014

Sigma-Aldrich

Collagenase from Clostridium histolyticum

0.2 μm filtered, Type V-S, ≥1 FALGPA units/mg solid, ≥125 CDU/mg solid

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

Quality Level

sterility

0.2 μm filtered

form

lyophilized powder

specific activity

≥1 FALGPA units/mg solid
≥125 CDU/mg solid

mol wt

68-125 kDa

storage temp.

−20°C

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This Item
C8176C8051C0773
form

lyophilized powder

form

powder

form

powder

form

lyophilized powder

specific activity

≥1 FALGPA units/mg solid, ≥125 CDU/mg solid

specific activity

≤1.0 FALGPA units/mg solid

specific activity

≥1.0 FALGPA units/mg solid

specific activity

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

mol wt

68-125 kDa

mol wt

68-130 kDa

mol wt

68-130 kDa

mol wt

68-130 kDa

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

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

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 (pH optimum: 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.

Preparation Note

Prepared from Type V (C9263)

Analysis Note

Also contains clostripain, nonspecific neutral protease, and tryptic activities.

substrate

Product No.
Description
Pricing

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

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[Dupuytren contracture].
R E Giunta et al.
MMW Fortschritte der Medizin, 153(49-50), 41-43 (2012-02-09)
Jörg Witthaut et al.
Clinical drug investigation, 31(11), 791-798 (2011-10-05)
Injectable collagenase Clostridium histolyticum is efficacious in correcting Dupuytren's contracture as assessed by changes in the angle of contracture and range of motion (ROM). However, clinically important changes in ROM have not been evaluated in depth. The objective of this
Skin laceration as a serious adverse sequela of injectable collagenase for Dupuytren contracture.
Geoffrey G Hallock
Plastic and reconstructive surgery, 129(1), 205e-206e (2011-12-22)
Chris Bainbridge et al.
Journal of plastic surgery and hand surgery, 46(3-4), 177-183 (2012-06-08)
Collagenase Clostridium histolyticum (CCH) is a non-surgical, efficacious therapy for Dupuytren's contracture (DC). This study evaluated the efficacy and safety of CCH in patients with previous DC surgery. Data from 12 CCH clinical trials were pooled. At screening, patients provided
Thomas J Edkins et al.
Clinical and vaccine immunology : CVI, 19(4), 562-569 (2012-02-24)
Collagenase Clostridium histolyticum (CCH) contains a fixed ratio of class I (AUX-I) and class II (AUX-II) collagenases and is used as treatment for Dupuytren's contracture. These two Zn-dependent enzymes, produced by the Gram-positive bacterium Clostridium histolyticum, are related functionally to

Protocols

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

To measure collagenase activity, N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala is used in a continuous spectrophotometric rate determination at 345 nm. Collagenase hydrolyzes collagen peptide bonds.

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