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C0255

Sigma-Aldrich

Collagenase from Clostridium histolyticum

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

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

form

lyophilized powder

Quality Level

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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This Item
C0773C2399C9697
specific activity

≥500 CDU/mg solid (CDU = collagen digestion units)

specific activity

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

specific activity

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

specific activity

≥800 units/mg solid

mol wt

68-130 kDa

mol wt

68-130 kDa

mol wt

-

mol wt

68-130 kDa

purified by

chromatography

purified by

chromatography

purified by

chromatography

purified by

-

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

-

foreign activity

Clostripain ≤2.0 U/mg

foreign activity

caseinase and clostripain ≤1 units/mg protein

foreign activity

-

foreign activity

-

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 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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Invasive trophoblast promote stromal fibroblast decidualization via Profilin 1 and ALOX5
Menkhorst EM, et al.
Scientific reports, 7(1), 8690-8690 (2017)
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
Solution structure of clostridial collagenase H and its calcium-dependent global conformation change
Ohbayashi N, et al.
Biophysical Journal, 104(7), 1538-1545 (2013)
Structural stability enhancement of full-length clostridial collagenase by calcium ions
Ohbayashi N, et al.
Applied and Environmental Microbiology, AEM-00808 (2012)
Electrophoretic cytopathology resolves ERBB2 forms with single-cell resolution
Kang CC, et al.
npj precision oncology, 2(1), 10-10 (2018)

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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