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C8051

Sigma-Aldrich

Collagenase from Clostridium histolyticum

Sigma Blend Type H, ≥1.0 FALGPA units/mg solid

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

Quality Level

form

powder

caseinase activity

≥10 units/mg solid

specific activity

≥1.0 FALGPA units/mg solid

mol wt

68-130 kDa

storage temp.

−20°C

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This Item
C8176C7926C2014
form

powder

form

powder

form

powder

form

lyophilized powder

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

mol wt

68-130 kDa

mol wt

68-130 kDa

mol wt

68-130 kDa

mol wt

68-125 kDa

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

caseinase activity

≥10 units/mg solid

caseinase activity

≥40 units/mg solid

caseinase activity

≤10 units/mg solid

caseinase activity

-

Application

Collagenase from Clostridium histolyticum has been used:
  • to harvest osteosarcoma 143B cells from collagen scaffold
  • in tissue digestion of islets and carotid artery
  • in the digestion of chorionic villi
  • in enzymatic dissociation for ATP-based tumour chemosensitivity assay (ATP-TCA) in solid tumors

Biochem/physiol Actions

Clostridium histolyticum produces two classes of collagenases. Clostridial collagenases are found to be highly effective than mammalian collagenases. It is capable of cleaving at multiple cleavage sites within the collagen triple helix. It is implicated in the 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. 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. Collagenase treatment can cause some cells to die. Typically, concentrations varying from 0.1 to 5 mg/mL is used for digestion. The duration of reaction varyies from 15 minutes to several hours to yield a satisfactory efficiency of cell dissociation, without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preffered and Zn2+ is required for activity. This enzyme is tested for the release of hepatocytes at approximately 1 mg/mL conc., in a total volume of 100 mL for each rat liver.
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.

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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Ming Yu et al.
Journal of applied physiology (Bethesda, Md. : 1985), 93(4), 1391-1399 (2002-09-18)
In contrast to its constrictor effects on peripheral arteries, 20-hydroxyeicosatetraenoic acid (20-HETE) is an endothelial-dependent dilator of pulmonary arteries (PAs). The present study examined the hypothesis that the vasodilator effects of 20-HETE in PAs are due to an elevation of
The in vitro effect of gefitinib (Iressa) alone and in combination with cytotoxic chemotherapy on human solid tumours
Knight LA, et al.
BMC Cancer, 4(1), 83-83 (2004)
Growth inhibition of osteosarcoma cell lines in 3D cultures: role of nitrosative and oxidative stress.
Gorska M, et al.
Anticancer Research, 36(1), 221-229 (2016)
Exendin-4 improves yield and function of isolated pre-weaned porcine islets
Lau H, et al.
Journal of Endocrinology and Diabetes, 5(5), 1-7 (2018)
Erik J Behringer et al.
British journal of pharmacology, 166(2), 774-787 (2011-12-16)
Electrical conduction along endothelium of resistance vessels has not been determined independently of the influence of smooth muscle, surrounding tissue or blood. Two interrelated hypotheses were tested: (i) Intercellular conduction of electrical signals is manifest in endothelial cell (EC) tubes;

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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