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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:
UNSPSC Code:
12352204
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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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 hazardExclamation mark

signalword

Danger

Hazard Classifications

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

target_organs

Respiratory system

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

dust mask type N95 (US), Eyeshields, Faceshields, Gloves


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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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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