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C2399

Sigma-Aldrich

Collagenase from Clostridium histolyticum

0.2 μm filtered, high purity, purified by chromatography, Type VII-S, ≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)

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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

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

purified by

chromatography

solubility

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

storage temp.

−20°C

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1 of 4

This Item
C0773C0255C5894
form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

solubility

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

solubility

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

solubility

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

solubility

-

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

sterility

0.2 μm filtered

sterility

-

sterility

-

sterility

non-sterile; 0.2 μm filtered

purified by

chromatography

purified by

chromatography

purified by

chromatography

purified by

-

Application

Collagenase from Clostridium histolyticum has been used:
  • to induce intracerebral hemorrhage (ICH) in mice
  • to digest collagen 3D gels for immunofluorescence
  • to cause focal blood extravasation and hemorrhage in mouse

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

Clostridium histolyticum contains two classes of collagenases. Clostridial collagenases are more effective than mammalian collagenases. It cleaves at multiple cleavage sites within the collagen triple helix. Clostridial collagenases are involved in bacterial invasion in gas gangrene.
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 VII (C0773). 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.

Substrate

Product No.
Description
Pricing

pictograms

Health hazard

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

Storage Class

11 - Combustible Solids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

dust mask type N95 (US), Eyeshields, Faceshields, Gloves


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Yasunori Asao et al.
Biochemical and biophysical research communications, 529(3), 590-595 (2020-08-02)
Intracerebral hemorrhage (ICH) is one of the most severe subtypes of stroke with high morbidity and mortality. Although a lot of drug discovery studies have been conducted, the drugs with satisfactory therapeutic effects for motor paralysis after ICH have yet
Nrg1 Intracellular Signaling Is Neuroprotective upon Stroke
Navarro-Gonzalez C, et al.
Oxidative Medicine and Cellular Longevity, 2019 (2019)
Manesh Chittezhath et al.
Experimental cell research, 314(19), 3551-3562 (2008-10-07)
The SYT proto-oncoprotein (also known as SS18) is a gene expression regulator conserved across species. Although its biological function is still unknown, the importance of SYT as a housekeeping protein is illustrated by the lethal phenotype of SYT-null embryos. Notably
Sandrine Bergeron et al.
Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism, 39(6), 1015-1025 (2018-01-16)
Cerebral microbleeds (CMBs) could contribute to cognitive impairment in the general population and in patients with dementia. We designed a study to (i) develop a murine model of CMBs, (ii) assess whether CMBs affect cognition in this model and (iii)
Charlotte Germonpré et al.
Epilepsy research, 164, 106364-106364 (2020-06-05)
Intracerebral hemorrhage (ICH) is a known risk factor for the development of seizures, but little is known about the pathophysiology of seizures in the acute phase post-ICH and their influence on functional outcome. With the use of an animal model

Protocols

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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