COLLH-RO

Collagenase H

from Clostridium histolyticum

• UNSPSC Code: 12352204
• EC Number: 3.4.24.3 (BRENDA, IUBMB)
• NACRES: NA.54

Properties

• Product Name: Collagenase H, from Clostridium histolyticum
• 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
• Quality Level: 100

Description

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.

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 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)

Safety Information

• pictograms: GHS07,GHS08
• signalword: Danger
• hcodes: H315 - H319 - H334 - H335
• pcodes: P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• target_organs: Respiratory system
• Storage Class: 11 - Combustible Solids
• wgk: WGK 1
• flash_point_f: does not flash
• flash_point_c: does not flash