COLLH-RO

Collagenase H

from Clostridium histolyticum

• Numéro de classification (Commission des enzymes): 3.4.24.3 (BRENDA, IUBMB)
• Code UNSPSC : 12352204

Propriétés

• Source biologique: Clostridium histolyticum
• Niveau de qualité: 100
• Stérilité: non-sterile
• Forme: lyophilized
• collagenase activity: 0.15 U/mg lyophilizate
• Conditionnement: pkg of 100 mg (11074032001); pkg of 2.5 g (11087789001); pkg of 500 mg (11074059001)
• Fabricant/nom de marque: Roche
• Concentration: ~1 mg/mL (Isolation of rat hepatocytes); ~2 mg/mL (Isolation of adipocytes)
• Technique(s): tissue processing: suitable
• Couleur: brown
• pH optimal: 6.0-8.0
• Solubilité: water: soluble
• Numéro d'accès NCBI: WP_138210331
• Numéro d'accès UniProt: Q46085
• Application(s): life science and biopharma
• Activité étrangère: Clostripain 39.798 U/mg; Proteases 14.014 U/mg (Azocoll, based on lyoph); Trypsin 1.668 U/mg (with BAEE)
• Température de stockage: 2-8°C

Description

Description générale

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

Spécificité

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.

Forme physique

Lyophilizate, nonsterile

Notes préparatoires

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)

Stockage et stabilité

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

Autres remarques

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

Informations sur la sécurité

• Pictogrammes: GHS07,GHS08
• Mention d'avertissement: Danger
• Mentions de danger: H315 - H319 - H334 - H335
• Conseils de prudence: P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311
• Classification des risques: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Organes cibles: Respiratory system
• Code de la classe de stockage: 11 - Combustible Solids
• Classe de danger pour l'eau (WGK): WGK 1
• Point d'éclair (°F): does not flash
• Point d'éclair (°C): does not flash