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COLLH-RO

Roche

Collagenase H

Name: Collagenase H
Brand: ROCHE

from Clostridium histolyticum

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UNSPSC Code:

12352204

EG-Nummer:

3.4.24.3 (BRENDA, IUBMB)

NACRES:

NA.54

Biological source:

Clostridium histolyticum

Concentration:

~1 mg/mL (Isolation of rat hepatocytes), ~2 mg/mL (Isolation of adipocytes)

Technischer Dienst

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Unterstützung erhalten

Eigenschaften

biological source

Clostridium histolyticum

sterility

non-sterile

form

lyophilized

collagenase activity

0.15 U/mg lyophilizate

packaging

pkg of 100 mg (11074032001), pkg of 2.5 g (11087789001), pkg of 500 mg (11074059001)

manufacturer/tradename

Roche

concentration

~1 mg/mL (Isolation of rat hepatocytes), ~2 mg/mL (Isolation of adipocytes)

technique(s)

tissue processing: suitable

color

brown

optimum pH

6.0-8.0

solubility

water: soluble

NCBI accession no.

WP_138210331

UniProt accession no.

Q46085

application(s)

life science and biopharma

foreign activity

Clostripain 39.798 U/mg, Proteases 14.014 U/mg (Azocoll, based on lyoph), Trypsin 1.668 U/mg (with BAEE)

storage temp.

2-8°C

Beschreibung

General description

Collagenase H is an enzyme mixture prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization.

Application

Collagenase from C. histolyticum is used for the dissociation of tissues for the establishment of primary cell cultures. Collagenase H is well suited for the isolation of hepatocytes from rat liver by the collagenase perfusion method. This enzyme preparation is also suitable for the preparation of other types of cells such as endothelial cells from large vessels, and for the isolation of adipocytes from epididymal fat pads of rats.

Biochem/physiol Actions

Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases also degrade other proteins.

Physical form

Lyophilizate, nonsterile

Preparation Note

Enzyme activity:
Collagenase activity: >0.15U/mg (according to Wünsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate).
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase H has a balanced ratio of enzyme activities, and is function tested for the isolation of rat hepatocytes (perfusion method).
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean)
Activator: Ca²⁺
Working concentration: 0.5 to 2.5mg/ml,
Approximately 1 mg/ml for the isolation of rat hepatocytes and 2mg/ml for the isolation of adipocytes.
Storage conditions (working solution): -15 to -25°C
The reconstituted solution should be stored at -15 to -25°C for short term storage, -60°C or as described below for long-term storage.

Keep container tightly closed in a dry and well-ventilated place.

Reconstitution in any balanced salt solution (e.g., HBSS)