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C0255

Collagenase aus Clostridium histolyticum

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

purified by chromatography, ≥500 CDU/mg solid (CDU = collagen digestion units), lyophilized powder

Synonym(e):

Clostridiopeptidase A

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Über diesen Artikel

CAS-Nummer:

9001-12-1

UNSPSC Code:

12352204

NACRES:

NA.54

EC Number:

232-582-9

MDL number:

MFCD00130830

EG-Nummer:

3.4.24.3 (BRENDA, IUBMB)

Specific activity:

≥500 CDU/mg solid (CDU = collagen digestion units)

Technischer Dienst

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Eigenschaften

form

lyophilized powder

Quality Segment

200

specific activity

≥500 CDU/mg solid (CDU = collagen digestion units)

mol wt

68-130 kDa

purified by

chromatography

solubility

TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C

foreign activity

Clostripain ≤2.0 U/mg

storage temp.

2-8°C

Beschreibung

General description

Collagenase from Clostridium histolyticum comprises of collagenases G and H. Collagenase has N-terminal activator domain and C-terminal peptidase domain. The collagen binding domains (CBD) and polycystic kidney disease-like (PKD-like) domain resides in the peptidase domain. The collagenase module undergoes structural changes upon calcium binding.

Application

Collagenase from Clostridium histolyticum has been used:

for the recovery of neutrophils from collagen gels
for the dissociation of breast tumor biopsies into single cells
for the isolation of primary human endometrial stromal cells(HESC) from endometrial biopsies

Collagenase from Clostridium histolyticum may be used for the isolation of cells from many types of animal tissue. It may be used for the disaggregation of human tumor, mouse kidney, human adult/fetal brain, lung and many other tissues. It may also be used in liver and kidney perfusion studies, for the digestion of pancreas, isolation of nonparenchymal rat liver cells and for hepatocyte preparation.

Biochem/physiol Actions

Collagenase wird durch vier Grammatom Calcium pro Mol Enzym aktiviert. Sie wird durch Ethylenglycol-bis-(beta-aminoethylether)-N, N, N′,N′-tetraessigsäure, β-Mercaptoethanol, Glutathion, Thioglycolsäure und 8-Hydroxychinolin gehemmt.

Crude collagenases are mixtures of enzymes (mostly proteases) secreted by C. histolyticum. 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.

Packaging

Product sold on the basis of collagen digestion units (CDU).

Preparation Note

Collagenase may be dissolved in TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37 °C) at 0.05-0.1 mg/mL concentration.

Other Notes

Eine Collagen-Aufschlusseinheit (Collagen Digestion Unit, CDU) setzt Peptide aus Rinder-Achillessehnencollagen entsprechend der Ninhydrin-Anfärbung von 1,0 μmol Leucin in 5 Stunden bei pH 7,4 und 37 °C in Gegenwart von Calciumionen frei. Eine FALGPA-Hydrolyseeinheit hydrolysiert 1,0 μmol Furylacryloyl-Leu-Gly-Pro-Ala pro Minute bei 25 °C. Eine neutrale Protease-Einheit hydrolysiert die Caseinmenge, die eine Färbung entsprechend 1,0 μmol Tyrosin pro 5 Stunden bei pH 7,5 und 37 °C produziert. Eine Clostripain-Einheit hydrolysiert 1,0 μmol BAEE pro Minute bei pH 7,6 und 25 °C in Gegenwart von DTT.