Not available outside of the UK & Ireland.

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.

General description

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

Other Notes

For life science research only. Not for use in diagnostic procedures.

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 activityCollagenase 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-mercaptoethanolCollagenase is not inhibited by serum.Clostripain inhibitors: TLCKTrypsin 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°CThe reconstituted solution should be stored at -15 to -25°C for short term storage, -60°C or as described below for long-term storage.

Reconstitution

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

Specificity

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