C9722

Colagenasa from Clostridium histolyticum

Name: Colagenasa from <I>Clostridium histolyticum</I>
Brand: SIGMA

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

Sinónimos:

Clostridiopeptidasa A

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About This Item

CAS Number:

9001-12-1

Número CE:

3.4.24.3 (BRENDA, IUBMB)

Número CE:

232-582-9

Número MDL:

MFCD00130830

Código UNSPSC:

12352204

NACRES:

NA.75

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origen biológico

Clostridium histolyticum

Nivel de calidad

200

esterilidad

0.2 μm filtered

Formulario

lyophilized powder (from 0.2μm filtered solution)

actividad específica

0.5-5.0 FALGPA units/mg solid

concentración

1 mg/mL

técnicas

cell culture | mammalian: suitable
single cell analysis: suitable

pH

7.4

Condiciones de envío

ambient

temp. de almacenamiento

−20°C

Aplicación

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.

Acciones bioquímicas o fisiológicas

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.

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.

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

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.

Nota de preparación

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.

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