C2014

Collagenase from Clostridium histolyticum

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

0.2 μm filtered, Type V-S, ≥1 FALGPA units/mg solid, ≥125 CDU/mg solid

Synonyma:

Clostridiopeptidase A

Přihlásitk zobrazení cen stanovených pro organizaci a smluvních cen

About This Item

CAS Number:

9001-12-1

Číslo EC:

3.4.24.3 (BRENDA, IUBMB)

EC Number:

232-582-9

MDL number:

MFCD00130830

UNSPSC Code:

12352204

NACRES:

NA.54

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Quality Level

200

sterility

0.2 μm filtered

form

lyophilized powder

specific activity

≥1 FALGPA units/mg solid
≥125 CDU/mg solid

mol wt

68-125 kDa

storage temp.

−20°C

Application

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.

Biochem/physiol Actions

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 recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid.

Unit Definition

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.