C0255

Sigma-Aldrich

Collagenase from Clostridium histolyticum

purified by chromatography, ≥0.25 FALGPA units/mg solid, ≥250 CDU/mg solid (CDU = collagen digestion units), lyophilized powder

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

Quality Level

sterility

sterile-filtered

form

lyophilized powder

specific activity

≥0.25 FALGPA units/mg solid
≥250 CDU/mg solid (CDU = collagen digestion units)

mol wt

68-130 kDa

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

foreign activity

Clostripain ≤2.0 U/mg

storage temp.

−20°C

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General description

Collagenase from Clostridium histolyticum comprises of collagenases G and H. Collagenase has N-terminal activator domain and C-terminal peptidase domain. The collagen binding domains (CBD) and polycystic kidney disease-like (PKD-like) domain resides in the peptidase domain. The collagenase module undergoes structural changes upon calcium binding.

Application

Collagenase from Clostridium histolyticum may be used for the isolation of cells from many types of animal tissue. It may be used for the disaggregation of human tumor, mouse kidney, human adult/fetal brain, lung and many other tissues. It may also be used in liver and kidney perfusion studies, for the digestion of pancreas, isolation of nonparenchymal rat liver cells and for hepatocyte preparation.
Collagenase from Clostridium histolyticum has been used:
  • for the recovery of neutrophils from collagen gels
  • for the dissociation of breast tumor biopsies into single cells
  • for the isolation of primary human endometrial stromal cells(HESC) from endometrial biopsies

Packaging

7500 units in poly bottle
1500, 3000 units in glass bottle
Product sold on the basis of collagen digestion units (CDU).

Biochem/physiol Actions

Crude collagenases are mixtures of enzymes (mostly proteases) secreted by C. histolyticum. 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.
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

Collagenase may be dissolved in TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37 °C) at 0.05-0.1 mg/mL concentration.

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

Solution structure of clostridial collagenase H and its calcium-dependent global conformation change
Ohbayashi N, et al.
Biophysical Journal, 104(7), 1538-1545 (2013)
V Pendaries et al.
Gene therapy, 17(7), 930-937 (2010-04-09)
Recessive dystrophic epidermolysis bullosa (RDEB) is a severe genodermatosis caused by loss-of-function mutations in COL7A1 encoding type VII collagen, the component of anchoring fibrils. As exogenous type VII collagen may elicit a deleterious immune response in RDEB patients during upcoming...
Structural stability enhancement of full-length clostridial collagenase by calcium ions
Ohbayashi N, et al.
Applied and Environmental Microbiology, AEM-00808 (2012)
E M Menkhorst et al.
Scientific reports, 7(1), 8690-8690 (2017-08-20)
During the establishment of pregnancy, extravillous trophoblast (EVT) must invade into the uterine decidua to facilitate decidual artery remodelling to create the placental blood supply. The local decidual environment is thought to regulate trophoblast invasion, however these interactions are poorly...
Electrophoretic cytopathology resolves ERBB2 forms with single-cell resolution
Kang CC, et al.
npj precision oncology, 2(1), 10-10 (2018)

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