C1889

Collagenase from Clostridium histolyticum

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

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

Synonim(y):

Clostridiopeptidase A

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About This Item

CAS Number:

9001-12-1

Numer WE:

3.4.24.3 (BRENDA, IUBMB)

Numer WE:

232-582-9

Numer MDL:

MFCD00130830

Kod UNSPSC:

12352204

NACRES:

NA.54

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Poziom jakości

200

sterylność

0.2 μm filtered

typ

Type IV-S

Formularz

powder

aktywność właściwa

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

masa cząsteczkowa

68-130 kDa

przydatność

suitable for release of physiologically active rat hepatocytes

Zastosowanie

diagnostic assay manufacturing

temp. przechowywania

−20°C

Opis ogólny

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.

Zastosowanie

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.

Działania biochem./fizjol.

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.

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.

Przestroga

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.

Definicja jednostki

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.

Uwaga dotycząca przygotowania

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.

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