C9407

Sigma-Aldrich

Collagenase from Clostridium histolyticum

powder, Suitable for the digestion and isolation of physiologically active pancreatic islet cells, suitable for cell culture

Synonym(s):
Clostridiopeptidase A
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.75

Quality Level

biological source

Clostridium histolyticum

form

powder

specific activity

≥800 units/mg solid

mol wt

68-130 kDa

packaging

pkg of 1 μmol

application(s)

cell culture | mammalian: suitable

pH

7.4

shipped in

dry ice

storage temp.

−20°C

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General description

Collagenase enzyme from Clostridium histolyticum has collagen binding domain (CBD) and polycystic kidney disease-like (PKD-like) domains. It also has Ca2+ binding site for calcium binding, which is crucial for its functionality. Collagenase is used in the management of connective tissue disorder, Dupuytren′s contracture.

Application

This product is suitable for the disaggregation of human tumor, mouse kidney, human adult and fetal brain, lung and many other epithelia tissues. It has also been shown to be effective in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocyte preparation. Collagenase has also been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products. This enzyme has been tested for the release of heptatocytes at a concentration of approximately 1 mg/mL. Concentrations for digestion range from 0.1 to 5 mg/mL.
Collagenase from Clostridium histolyticum has been used in the digestion:
  • of pancreatic samples for islets isolation
  • of pulmonary artery samples for pulmonary vascular smooth muscle cells (PVSMCs) isolation
  • of cartilage samples for chondrocytes isolation

Biochem/physiol Actions

The collagenase product is a mixture of enzymes secreted by C. histolyticum, with different products differentiated by the relative ratios of the 10-18 components found in the secreted enzymes. The main components are two collagenases, clostripain, and a neutral protease. The synergistic action of these enzymes degrade collagen and other intracellular materialThe action of both collagenase enzymes and the neutral protease is necessary for effective release of cells from tissue. Various types of collagen are the natural substrates for collagenase.
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.

Caution

As supplied, this product is stable for one year at -20°C. There is no loss in FALGPA or protease activity in 30 days at 37°C, 50°C and -20°C. Solutions of crude collagenase are stable if frozen quickly in aliquots (at 10 mg/mL) and kept frozen at -20°C. Further freeze-thaw cycles will damage the solution. The product retains 100% activity over 7 hours when held on ice.

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

This proudct is prepared from Type XI (C7657) Collagenase from Clostridium histolyticum. It also contains clostripain, nonspecific neutral protease and tryptic activities. Solutions are typically prepared at 1-2 mg/mL in TESCA buffer (containing 50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37°C.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

hazcat

Resp. Sens. 1

storage_class_code

11 - Combustible Solids

WGK Germany

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

Certificate of Origin

Phenotypic modulation of human articular chondrocytes by bistratene A
Gargiulo BJ, et al.
European Cells & Materials, 3(2), 9-18 (2002)
Endothelin receptor antagonists attenuate the inflammatory response of human pulmonary vascular smooth muscle cells to bacterial endotoxin
Knobloch J, et al.
Journal of Pharmacology and Experimental Therapeutics, 346(2), 290-299 (2013)
Y F Li et al.
Neoplasma, 67(2), 430-437 (2020-01-25)
Patient-derived organoids (PDOs) are emerging as preclinical models with promising values in personalized cancer therapy. The purpose of this study was to establish a living biobank of PDOs from patients with non-small cell lung cancer (NSCLC) and to study the...
Collagenase clostridium histolyticum in Dupuytren's contracture: a systematic review
Smeraglia F, et al.
British Medical Bulletin, 118(1), 149-149 (2016)
The cytoprotection of chitosan based hydrogels in xenogeneic islet transplantation: An in vivo study in streptozotocin-induced diabetic mouse
Yang KC, et al.
Biochemical and Biophysical Research Communications, 393(4), 818-823 (2010)

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