- Manufactured for superior lot-to-lot enzyme consistency for improved performance consistency
- Low contaminating endotoxin levels (<50.0 EU/mg)
- Lot specific Certificate of Analysis provided with complete QC analysis results
Cat. # 004-1000
CIzyme Clostripain is an aseptically dispensed, lyophilized purified unactivated protein from Clostidium histolyticum.
Sold in 25 mg bottles.
- White lyophilized cake under vacuum in amber bottle sealed with butyl rubber stopper
- Contains approximately 375,000 CDA units and 75,000 NPA units.
- Unopened product stable for at least 2 years when stored between -15 to -25°C; internal studies indicate comparable stability for reconstituted, frozen solution for at least 3 months at the same temperatures
- Shipped on dry ice to ensure enzyme stability during initial reconstitution.
This product is purified from culture supernatants of C. histolyticum cultures that contain porcine gelatin or porcine pancreatic enzyme derived from US and Canadian sources. No bovine derived animal products are used in any step used to prepare the purified collagenase enzymes.
CIzyme Clostripain is dispensed by mass and the total BAEE units of enzyme activities are detailed on the lot specific certificate of analysis. Treatment of a reducing agent will convert the unactivated form into the active form, which is approximately 10-fold higher by BAEE substrate.
The peptide substrate N-benzoyl-L-arginine ethyl ester (BAEE) is the traditional method of assaying clostripain activity. This peptide was originally developed to detect trypsin activity, but works for clostripain as well. The assay is run in the buffer described in the reference for the unactivated activity and also in the presence of a reducing agent (DTT) to determine the activated clostripain activity.
No references on file.
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 Protease FAQ
Yes. Selection of a protease in many cases is simply due to the historical precedence of the given cell isolation protocol being followed; however, that is not to say a different one would not work. The isolation of human islets is a perfect example in which thermolysin, BP Protease (a Dispase equivalent), and C. histolyticum neutral protease have all been successfully used. Generally, bacterial or fungal sources of protease are selected due to their broader specificity. Substitution of one protease for another may not be a seamless transition, however, as the kinetics of digestion can be dramatically different among the various general proteases. For guidance in selecting a protease, contact VitaCyte’s technical support team.
This product is stable for at least two years from date of manufacture if stored unopened between -15 to – 25°C. Internal studies have shown the reconstituted enzyme is stable as a frozen solution between -15 to – 25°C for at least 6 months without loss of potency. Additional internal studies have shown the enzyme can go through a freeze-thaw cycle at least once without loss of potency by neutral protease (NP) assay. The product is shipped on dry ice to provide the most stable conditions during shipment.
No. Advances in cell transplantation procedures may lead to the use of this product to prepare cells for clinical transplantation. The guidance for use of research reagents in these procedures is governed by existing laws and regulations. This includes approval of human experimental protocols by a local Institutional Review Board. Responsibility for applying this product in any clinical procedure lies solely with the Principal Investigator and Institution supervising this activity. If you plan to use this product in any clinical transplant procedure, please consult the policies of your institution or those found on the National Institutes of Health website: http://www.hhs.gov/ohrp/