Collagenase from Clostridium histolyticum
SIGMA/C2799 - powder, suitable for cell culture, ≥4 FALGPA units/mg solid, high purity, ≥700 CDU/mg solid (CDU = collagen digestion units)
Synonym: Clostridiopeptidase A
CAS Number: 9001-12-1
EC Number: 232-582-9
MDL Number: MFCD00130830
Product Type: Chemical
Sigma collagenases are fully characterized by enzymatic activity. Many are specifically tested for suitability for cell dissociation and cell viability for hepatocytes, adipocytes, and pancreatic cells.
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Unit Definition One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0 μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions.
| biological source | Clostridium histolyticum |
| foreign activity | neutral protease and clostripain ≤1 unit/mg solid |
| form | powder |
| mol wt | 68-130 kDa |
| pH | 7.4 |
| purified by | chromatography |
| Quality Level | 200  |
| shipped in | dry ice |
| specific activity | ≥4 FALGPA units/mg solid |
| ≥700 CDU/mg solid (CDU = collagen digestion units) | |
| storage temp. | −20°C |
| technique(s) | cell culture | mammalian: suitable |
| Application: | This product is suitable for the disaggregation of human tumor, mouse kidney, human adult and fetal brain, lung and many other epithelia tissues. It has also been shown to be effective in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocyte preparation. Collagenase has also been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products. This enzyme has been tested for the release of heptatocytes at a concentration of approximately 1 mg/mL. Concentrations for digestion range from 0.1 to 5 mg/mL. |
| Application: | Use for the digestion of collagen and the release of adherent cells from substrates. |
| Biochem/physiol Actions: | Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(beta-aminoethy |
| Biochem/physiol Actions: | The collagenase product is a mixture of enzymes secreted by C. histolyticum, with different products differentiated by the relative ratios of the 10-18 components found in the secreted enzymes. The main components are two collagenases, clostripain, and a neutral protease. The synergistic action of these enzymes degrade collagen and other intracellular material. The action of both collagenase enzymes and the neutral protease is necessary for effective release of cells from tissue. Various types of collagen are the natural substrates for collagenase. |
| Disclaimer: | As supplied, this product is stable for one year at -20°C. There is no loss in FALGPA or protease activity in 30 days at 37°C, 50°C and -20°C. Solutions of crude collagenase are stable if frozen quickly in aliquots (at 10 mg/mL) and kept frozen at -20°C. Further freeze-thaw cycles will damage the solution. The product retains 100% activity over 7 hours when held on ice. |
| Other Notes: | One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0 μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions. One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of furylacryloyl-Leu-Gly-Pro |
| Preparation Note: | This collagenase is obtained from the culture filtrate of type VII (C0773) Clostridium histolyticum. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. Solutions are typically prepared at 1-2 mg/mL in TESCA buffer (containing 50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37°C. |
| Symbol |  GHS08 |
| Signal word | Danger |
| Hazard statements | H334 |
| Precautionary statements | P261 - P284 - P501 |
| Hazard Codes | Xn |
| Risk Statements | 36/37/38-42 |
| Safety Statements | 22-24-26-36/37 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| activity | specific activity: ≥4 FALGPA units/mg solid; specific activity: ≥700 CDU/mg solid (CDU = collagen digestion units) |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 3.4.24.3 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |