C9697

Collagenasi

lyophilized powder (from 0.2 μm filtered solution), suitable for cell culture

• Sinonimo/i: Clostridiopeptidasi A
• CAS Number: 9001-12-1
• Numero CE: 3.4.24.3 (BRENDA, IUBMB)
• Numero CE: 232-582-9
• Numero MDL: MFCD00130830
• Codice UNSPSC: 12352204
• NACRES: NA.54

Proprietà

• Origine biologica: Clostridium histolyticum
• Livello qualitativo: 200
• Sterilità: 0.2 μm filtered
• Stato: lyophilized powder (from 0.2 μm filtered solution)
• Attività specifica: ≥800 units/mg solid
• PM: 68-130 kDa
• Concentrazione: 1 mg/mL
• tecniche: cell culture | mammalian: suitable
• pH: 7.4
• Temperatura di conservazione: −20°C

Descrizione

Descrizione generale

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.

Applicazioni

Collagenase from Clostridium histolyticum is used for the following applications:

• 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.

Azioni biochim/fisiol

La collagenasi è attivata da quattro grammi atomo di calcio per mole di enzima. È inibita da etilen-glicol-bis(beta-amminoetil etere) - acido N, N, N′,N′-tetraacetico, beta-mercaptoetanolo, glutatione, acido tioglicolico e 8-idrossichinolina.

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.

Avvertenza

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.

Definizione di unità

Una unità di digestione del collagene (CDU) libera peptidi da collagene proveniente da tendine d′Achille bovino equivalente in colore ninidrina a 1,0 μmole di leucina in 5 ore a pH 7,4 a 37 °C in presenza di ioni calcio. Una unità di idrolisi FALGPA idrolizza 1,0 μmole di furilacriloil-Leu-Gly-Pro-Ala per minuto a 25 °C. Una unità di proteasi neutra idrolizza caseina e produce colore equivalente a 1,0 μmole di tirosina per 5 ore a pH 7,5 a 37 °C. Una unità di clostripaina idrolizza 1,0 μmole of BAEE per minuto a pH 7,6 a 25 °C in presenza di DTT.

Nota sulla preparazione

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).

Informazioni sulla sicurezza

• Pittogrammi: GHS08,GHS07
• Avvertenze: Danger
• Indicazioni di pericolo: H315,H319,H334,H335
• Consigli di prudenza: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Classi di pericolo: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Organi bersaglio: Respiratory system
• Codice della classe di stoccaggio: 11 - Combustible Solids
• Classe di pericolosità dell'acqua (WGK): WGK 1
• Punto d’infiammabilità (°F): Not applicable
• Punto d’infiammabilità (°C): Not applicable
• Dispositivi di protezione individuale: dust mask type N95 (US), Eyeshields, Faceshields, Gloves