C1889

Colagenasa from Clostridium histolyticum

0.2 μm filtered, suitable for release of physiologically active rat hepatocytes, Type IV-S, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid

• Sinónimos: Clostridiopeptidasa A
• Número de CAS: 9001-12-1
• Comisión internacional de enzimas: 3.4.24.3 (BRENDA, IUBMB)
• EC Number: 232-582-9
• MDL number: MFCD00130830
• UNSPSC Code: 12352204
• NACRES: NA.54

Propiedades

• Quality Level: 200
• sterility: 0.2 μm filtered
• type: Type IV-S
• form: powder
• specific activity: ≥125 CDU/mg solid; 0.5-5.0 FALGPA units/mg solid
• mol wt: 68-130 kDa
• suitability: suitable for release of physiologically active rat hepatocytes
• application(s): diagnostic assay manufacturing
• storage temp.: −20°C

Descripción

General description

Clostridium histolyticum consists of two classes of collagenases, which cleaves at multiple cleavage sites within the collagen triple helix. It is more effective than mammalian collagenases. Clostridial collagenase is implicated in bacterial invasion in gas gangrene.

Application

Collagenase from Clostridium histolyticum has been used to dissociate tissue samples such as spleens and testicular tissue.

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. When this enzyme is tested for suitability for the release of hepatocytes. The collagenase is used at approximately 1 mg/mL in a total volume of 100 mL for each rat liver.

Biochem/physiol Actions

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 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 varies from 15 minutes to several hours, which enables efficient cell dissociation without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preffered and Zn²⁺ is required for its activity.

La colagenasa es activada por cuatro átomo-gramos de calcio por mol de enzima. Es inhibida por el ácido etilenglicol-bis(beta-aminoetil éter) - N, N, N′,N′-tetraacético, el beta-mercaptoetanol, el glutatión, el ácido tioglicólico y la 8-hidroxiquinolina.

Caution

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.

Unit Definition

Una unidad de digestión de colágeno (CDU) libera péptidos del colágeno del tendón de Aquiles bovino que equivalen en la reacción de color de la ninhidrina a 1,0 μmoles de leucina en 5 horas a pH 7,4 y 37 °C en presencia de iones calcio. Una unidad de hidrólisis FALGPA hidroliza 1,0 μmoles de furilacriloil-Leu-Gly-Pro-Ala por minuto a 25°C. Una unidad de proteasa neutra hidroliza la caseína para producir color equivalente a 1,0 μmoles de tirosina por 5 horas a pH 7,5 y 37°C. Una unidad de clostripaína hidroliza 1,0 μmoles de BAEE por minuto a pH 7,6 y 25°C en presencia de DTT.

Preparation Note

Prepared from Type IV (C5138). A stock solution may be prepared by dissolving 0.05-0.1 mg/mL of collagenase in 50 mM TES buffer containing 0.36 mM calcium chloride (TESCA) (pH 7.4) at 37 °C. This product also contains clostripain, nonspecific neutral protease, and tryptic activities.

Información de seguridad

• pictograms: GHS08,GHS07
• signalword: Danger
• hcodes: H315,H319,H334,H335
• pcodes: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• target_organs: Respiratory system
• Storage Class: 11 - Combustible Solids
• wgk_germany: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable
• ppe: dust mask type N95 (US), Eyeshields, Faceshields, Gloves