- Approximately 85-90% of protein recovered from the enrichment procedure is collagenase
- Purification process eliminates the majority of contaminating protease activity including clostripain (trypsin-like activity)
- Weigh out what you need or reconstitute and use the entire bottle
- Costs 33-50% less than purified enzymes typically used for human islet isolation
- Combine with your choice of neutral protease for your established protocol
- Lot specific Certificate of Analysis provided with complete QC analysis results
Collagenase Gold is low protease, enriched collagenase product from Clostridium histolyticum that can be used for isolating cells from tissue or recovering adherent cells after in vitro culture. The product has been used successfully for human islet isolation. This material contains a high percentage of intact class I collagenase, a factor shown to contribute to higher human islet yields.
- White lyophilized cake under vacuum in amber bottle sealed with butyl rubber stopper
- Each bottle contains a minimally hygroscopic, polypeptide excipient that enables easy weighing for intended use and protects the product from enzyme degradation during storage
Collagenase Gold is an aseptically filled, lyophilized preparation of enriched Clostridium histolyticum collagenase. Approximately 85-90% of the C. histolyticum protein in this preparation is collagenase. Collagenase is obtained from culture supernatants of C. histolyticum that contain porcine gelatin and pancreatic enzymes sterilized prior to bacterial fermentation. Collagenase Gold is labelled with a number that reflects the minimal amount of total collagenase FALGPA activity1 per bottle. Each bottle also contains a minimally hygroscopic, polypeptide excipient that enables the user to weigh out the amount of product required for use and also protects the product from enzyme degradation during storage. The product is sold as 100 mg (#011-1160) or 1g (#011-1060) pack sizes where the mass represents the dry weight of the protein.
Collagenase Gold is prepared for research use only.
Each lot of product is characterized for collagenase activity using the FALGPA peptide substrate1. The clostripain and trypsin-like activities are determined on the specific lot of enriched collagenase used to prepare the Collagenase Gold product2. The amount of these activities is calculated based on the amount of collagenase dispensed into each product.
Van Wart HE and Steinbrink DR. Analytical Biochemistry 113 (1981); 356-65.
Mitchell WM and Harrington WF. Methods in Enzymology 19 (1970) 635-642.
General Product FAQ
Thermolysin and BP Protease are entirely animal origin free.
A porcine gelatin peptone is used during the fermentation step of natural collagenase production. While this animal sourced material is required to stimulate collagenase production from the biosynthesis from the organism Clostridium histolyticum during anaerobic fermentation, it represents a very low adventitious agent risk as the media is sterilized at 121°C for 30 minutes prior to inoculation, which has been shown to inactivate many viruses.
The gelatin is removed during the purification process and no animal sourced materials are used in the remainder of the process.
Additional manufacturing details are available in product insert literature for each product.
There are two components to inactivation of the digestive enzymes commonly used for tissue dissociation. To inactivate protease activity, a serum-containing media or buffer is most effective. Buffers or media containing human serum albumin (HSA) are ineffective in suppressing protease activity, even at very high concentrations of protein (>10%). However, serum-containing buffers do not inhibit collagenase activity. The most effective method for inactivating collagenase activity is reducing temperature. Collagenases will lose about 80% their maximal activity at 26°C. Thus, introduction of an ice-cold, serum- containing media should virtually eliminate all collagenolytic and proteolytic activity of an enzyme mixture.
General Collagenase FAQ
VitaCyte’s technical service team has a significant amount of experience in transitioning from crude enzyme products to the more defined purified ones and is glad to provide you with assistance. If you can provide us details of the cell of interest; the enzyme product that is currently being used for the isolation; the concentrations (or activities) being targeted; volumes utilized; and, a certificate of analysis for the particular lot you would like to mimic; we can provide some reasonable guidance to which product(s) and what target activities best fit your application. Our defined enriched (DE) collagenase products are specifically designed to encompass the broad range of activities found in the crude enzyme products and to make the transition to a highly purified product seamless.
In developing an Application Specific Formulation, we generally rely on individuals considered “expert in the art” to ß-test potential formulations and guide us in the refinement of the final composition. These formulations are usually engineered from existing protocols and crude enzyme targets. Once a formulation has been identified and deemed acceptable by at least a couple independent labs, it may be included in our product portfolio. We are always eager to assist you with virtually any request for a custom blend so do not hesitate to contact us to discuss your application.