Crude collagenase preparations contain several isoforms of two different collagenases, a sulfhydryl protease, clostripain, a trypsin-like enzyme, and an aminopeptidase. This combination of collagenolytic and proteolytic activities is effective at breaking down intercellular matrices, the essential part of tissue dissociation. One component of the complex is a hydrolytic enzyme which degrades the helical regions in native collagen preferentially at the Y-Gly bond in the sequence Pro-Y-Gly-Pro, where Y is most frequently a neutral amino acid. This cleavage yields products susceptible to further peptidase digestion. Crude collagenase is inhibited by metal chelating agents such as cysteine, EDTA or o-phenanthroline but not DFP. It is also inhibited by α2-macroglobulin, a large plasma glycoprotein. Ca2+ is required for enzyme activity. Particular enzymatic profiles of each collagenase have been correlated with the tissues from which the cells for study were obtained (or with the uses to which the cells are put) and as a result of the correlations several types of crude collagenases have been established by We: Types 1, 2, 3, and 4.Type 1 crude collagenase has the original balance of collagenase, caseinase, clostripain and tryptic activities.Type 2 contains higher relative levels of protease activity, particularly clostripain. Type 3 contains lowest levels of secondary proteases. Type 4 is designed to be especially low in tryptic activity to limit damage to membrane proteins and receptors. Purified collagenase,, contains minimal secondary proteolytic activities along with high collagenase activity.NEW! Animal Origin Free Types AFA, AFB and AFC, STZ1, and STZ2 collagenases are derived from cultures grown in medium completely devoid of animal based components and designed for bioprocessing applications where introduction of potential animal derived pathogens must be prevented. Levels of secondary proteases are similar to Types 1 and 2 collagenase.CLSAFA is the original AFA grade designed to have collagenase and secondary proteases similar to Types 1 and 2 collagenase.CLSAFB contains higher collagenase and caseinase activities than CLSAFA.CLSAFC has especially low tryptic activity similar to Type 4 collagenase.STZ1 & STZ2, 0.22μ filtered STEMxyme AOF Collagenase/Neutral Protease (Dispase) blends for primary and stem cell isolation.
| Source | Clostridium histolyticum |
| Description | The original balance of enzymatic activities. Each lot assayed for collagenase, caseinase, clostripain and tryptic activities. Suggested for epithelial, liver, lung and adrenal primary cell isolations. A dialyzed, lyophilized powder. |
| Storage | Store at 2-8°C. |
| Activity | ≥125 units per mg dry weight |
| Unit Definition | One Unit releases one micromole of L-leucine equivalents from collagen in 5 hours at 37°C, pH 7.5 |
