C1889

Sigma-Aldrich

Collagenase from Clostridium histolyticum

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

Sinónimos:
Clostridiopeptidase A
Número de CAS:
Comisión internacional de enzimas:
Número de EC:
Número MDL:
NACRES:
NA.54

Nivel de calidad

200

esterilidad

0.2 μm filtered

formulario

powder

specific activity

≥125 CDU/mg solid
0.5-5.0 FALGPA units/mg solid

mol peso

68-130 kDa

idoneidad

release of physiologically active rat hepatocytes tested

Industria destacada

Diagnostic Assay Manufacturing

temp. de almacenamiento

−20°C

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Descripción general

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.

Aplicación

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.

Acciones bioquímicas o fisiológicas

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.
Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(beta-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, beta-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline.

Precaución

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.

Definición de unidad

One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0 μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions. One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of furylacryloyl-Leu-Gly-Pro-Ala per min at 25°C. One Neutral Protease unit hydrolyzes casein to produce color equivalent to 1.0 μmole of tyrosine per 5 hr at pH 7.5 at 37°C. One Clostripain Unit hydrolyzes 1.0 μmole of BAEE per min at pH 7.6 at 25°C in the presence of DTT.

Nota de preparación

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.

pictogramas

Health hazard

Palabra de señalización

Danger

Frases de peligro

Consejos de prudencia

hazcat

Resp. Sens. 1

storage_class_code

11 - Combustible Solids

WGK Alemania

WGK 1

Punto de inflamabilidad F

Not applicable

Punto de inflamabilidad C

Not applicable

Equipo de protección personal

dust mask type N95 (US), Eyeshields, Faceshields, Gloves

Certificado de Análisis

Certificado de origen

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