COLLH-RO

Collagenase H

from Clostridium histolyticum

• Enzyme Commission number: 3.4.24.3 (BRENDA, IUBMB)
• UNSPSC Code: 12352204

Properties

• biological source: Clostridium histolyticum
• Quality Level: 100
• 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

Description

General description

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

Specificity

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

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.

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.

Reconstitution

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

Storage and Stability

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

Other Notes

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

Safety Information

• Pictograms: GHS07,GHS08
• Signal Word: Danger
• Hazard Statements: H315 - H319 - H334 - H335
• Precautionary Statements: 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 Code: 11 - Combustible Solids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash