C2399

Collagenase aus Clostridium histolyticum

0.2 μm filtered, high purity, purified by chromatography, Type VII-S, ≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)

• Synonym(e): Clostridiopeptidase A
• CAS-Nummer: 9001-12-1
• EC-Nummer: 3.4.24.3 (BRENDA, IUBMB)
• EG-Nummer: 232-582-9
• MDL-Nummer: MFCD00130830
• UNSPSC-Code: 12352204
• NACRES: NA.54

Eigenschaften

• Qualitätsniveau: 200
• Sterilität: 0.2 μm filtered
• Form: lyophilized powder
• Spezifische Aktivität: ≥4 FALGPA units/mg solid; ≥700 CDU/mg solid (CDU = collagen digestion units)
• Aufgereinigt durch: chromatography
• Löslichkeit: TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C
• Lagertemp.: −20°C

Beschreibung

Anwendung

Collagenase from Clostridium histolyticum has been used:

• to induce intracerebral hemorrhage (ICH) in mice
• to digest collagen 3D gels for immunofluorescence
• to cause focal blood extravasation and hemorrhage in mouse

Collagenase has been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products (AGE). The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues. It is also effective in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes.

Biochem./physiol. Wirkung

Clostridium histolyticum contains two classes of collagenases. Clostridial collagenases are more effective than mammalian collagenases. It cleaves at multiple cleavage sites within the collagen triple helix. Clostridial collagenases are involved in 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 (pH optimum: 6.3-8.8). The enzyme recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid.

Collagenase wird durch vier Grammatom Calcium pro Mol Enzym aktiviert. Sie wird durch Ethylenglycol-bis-(beta-aminoethylether)-N, N, N′,N′-tetraessigsäure, β-Mercaptoethanol, Glutathion, Thioglycolsäure und 8-Hydroxychinolin gehemmt.

Einheitendefinition

Eine Collagen-Aufschlusseinheit (Collagen Digestion Unit, CDU) setzt Peptide aus Rinder-Achillessehnencollagen entsprechend der Ninhydrin-Anfärbung von 1,0 μmol Leucin in 5 Stunden bei pH 7,4 und 37 °C in Gegenwart von Calciumionen frei. Eine FALGPA-Hydrolyseeinheit hydrolysiert 1,0 μmol Furylacryloyl-Leu-Gly-Pro-Ala pro Minute bei 25 °C. Eine neutrale Protease-Einheit hydrolysiert die Caseinmenge, die eine Färbung entsprechend 1,0 μmol Tyrosin pro 5 Stunden bei pH 7,5 und 37 °C produziert. Eine Clostripain-Einheit hydrolysiert 1,0 μmol BAEE pro Minute bei pH 7,6 und 25 °C in Gegenwart von DTT.

Angaben zur Herstellung

Prepared from Type VII (C0773). A stock solution may be prepared by dissolving 0.05-0.1 mg of collagenase in one mL of 50 mM TES buffer containing 0.36 mM calcium chloride (TESCA), pH 7.4 at 37 °C.

Sicherheitshinweise

• Piktogramme: GHS08,GHS07
• Signalwort: Danger
• H-Sätze: H315,H319,H334,H335
• P-Sätze: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Gefahreneinstufungen: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Zielorgane: Respiratory system
• Lagerklassenschlüssel: 11 - Combustible Solids
• WGK: WGK 1
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable
• Persönliche Schutzausrüstung: dust mask type N95 (US), Eyeshields, Faceshields, Gloves