C9697
Collagenase aus Clostridium histolyticum
lyophilized powder (from 0.2 μm filtered solution), suitable for cell culture
Synonym(e):
Clostridiopeptidase A
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About This Item
Empfohlene Produkte
Biologische Quelle
Clostridium histolyticum
Qualitätsniveau
Sterilität
0.2 μm filtered
Form
lyophilized powder (from 0.2 μm filtered solution)
Spezifische Aktivität
≥800 units/mg solid
Mol-Gew.
68-130 kDa
Konzentration
1 mg/mL
Methode(n)
cell culture | mammalian: suitable
pH-Wert
7.4
Lagertemp.
−20°C
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Allgemeine Beschreibung
Collagenase is a protease which cleaves the triple-helical protein called collagen. There are three types of tissue collagenases, and these belong to the matrix metalloproteinases (MMP) family. Collagenase obtained from Clostridium histolyticum has a very strong activity, as it digests collagen from both ends, at temperatures as low as 4-10°C. Crude collagenase mixtures contain two major enzyme types namely, collagenase and clostripain.
Anwendung
Collagenase from Clostridium histolyticum is used for the following applications:
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 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 1mg/mL. Concentrations for digestion range from 0.1 to 5mg/mL.
- Sertoli cell isolation
- Used in the comparison of enzymatic methods
- Tissue preparations for immunocytochemistry
- Testicular sperm extraction
- Preparation of single cell suspensions
- Immunofluorescence
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 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 1mg/mL. Concentrations for digestion range from 0.1 to 5mg/mL.
Biochem./physiol. Wirkung
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.
Vorsicht
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.
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
Solutions are prepared from type XI at 1-2 mg/mL in TESCA buffer (containing 50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37°C).
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Signalwort
Danger
H-Sätze
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
Analysenzertifikate (COA)
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Protokolle
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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