C9722

Collagenase aus Clostridium histolyticum

lyophilized powder (from 0.2μm filtered solution), 0.5-5.0 FALGPA units/mg solid, suitable for cell culture

• Synonym(e): Clostridiopeptidase A
• CAS-Nummer: 9001-12-1
• UNSPSC Code: 12352204
• NACRES: NA.75
• EC Number: 232-582-9
• MDL number: MFCD00130830
• EG-Nummer: 3.4.24.3 (BRENDA, IUBMB)
• Specific activity: 0.5-5.0 FALGPA units/mg solid
• Biological source: Clostridium histolyticum
• Concentration: 1 mg/mL

Eigenschaften

• biological source: Clostridium histolyticum
• Quality Segment: 200
• sterility: 0.2 μm filtered
• form: lyophilized powder (from 0.2μm filtered solution)
• specific activity: 0.5-5.0 FALGPA units/mg solid
• concentration: 1 mg/mL
• technique(s): cell culture | mammalian: suitable, single cell analysis: suitable
• pH: 7.4
• shipped in: ambient
• storage temp.: −20°C

Beschreibung

Application

This product is suitable for the disaggregation of human tumor, mouse kidney, human adult and fetal brain, lung and many other epithelia tissues. It has also been shown to be effective in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocyte preparation. Collagenase has also been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products. This enzyme has been tested for the release of heptatocytes at a concentration of approximately 1 mg/mL. Concentrations for digestion range from 0.1 to 5 mg/mL.

Suitable for use in preparation of single cell suspension for sequencing.

Biochem/physiol Actions

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.

The collagenase product is a mixture of enzymes secreted by C. histolyticum, with different products differentiated by the relative ratios of the 10-18 components found in the secreted enzymes. The main components are two collagenases, clostripain, and a neutral protease. The synergistic action of these enzymes degrade collagen and other intracellular materialThe action of both collagenase enzymes and the neutral protease is necessary for effective release of cells from tissue. Various types of collagen are the natural substrates for collagenase.

Preparation Note

Solutions are typically prepared at 1-2 mg/mL in TESCA buffer (containing 50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37°C. This product also contains clostripain, nonspecific neutral protease and tryptic activities.

Other Notes

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.

Disclaimer

As supplied, this product is stable for one year at -20°C. There is no loss in FALGPA or protease activity in 30 days at 37°C, 50°C and -20°C. Solutions of crude collagenase are stable if frozen quickly in aliquots (at 10 mg/mL) and kept frozen at -20°C. Further freeze-thaw cycles will damage the solution. The product retains 100% activity over 7 hours when held on ice.

Sicherheitshinweise

• Piktogramme: GHS08,GHS07
• Signalwort: Danger
• Gefahrencodes: H315,H319,H334,H335
• Sicherheitshinweise: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Zielorgane: Respiratory system
• Lagerklasse: 11 - Combustible Solids
• Was ist los?: WGK 1
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable
• PSA (Persönliche Schutzausrüstung): dust mask type N95 (US), Eyeshields, Faceshields, Gloves