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C0255

Sigma-Aldrich

Collagenase from Clostridium histolyticum

purified by chromatography, ≥500 CDU/mg solid (CDU = collagen digestion units), lyophilized powder

Synonym(s):

Clostridiopeptidase A

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About This Item

CAS Number:
9001-12-1
Enzyme Commission number:
3.4.24.3 (BRENDA, IUBMB)
EC Number:
232-582-9
MDL number:
MFCD00130830
UNSPSC Code:
12352204
NACRES:
NA.54

form

lyophilized powder

Quality Level

200

specific activity

≥500 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 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

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.

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.

Packaging

Product sold on the basis of collagen digestion units (CDU).

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.

substrate

Product No.
Description
Pricing

Pictograms

Health hazardExclamation mark
GHS08,GHS07

Signal Word

Danger

Hazard Statements

H315,H319,H334,H335

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

Target Organs

Respiratory system

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Regulatory Information

常规特殊物品

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Structural stability enhancement of full-length clostridial collagenase by calcium ions
Ohbayashi N, et al.
Applied and Environmental Microbiology, AEM-00808 (2012)
Invasive trophoblast promote stromal fibroblast decidualization via Profilin 1 and ALOX5
Menkhorst EM, et al.
Scientific reports, 7(1), 8690-8690 (2017)
Electrophoretic cytopathology resolves ERBB2 forms with single-cell resolution
Kang CC, et al.
npj precision oncology, 2(1), 10-10 (2018)
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
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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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