Collagenase from Clostridium histolyticum
SIGMA/C7657 - Type XI, 2-5 FALGPA units/mg solid, ≥800 CDU/mg solid
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.
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.
| application(s) | diagnostic assay manufacturing |
| biological source | Clostridium histolyticum |
| form | powder |
| mol wt | 68-130 kDa |
| Quality Level | 200  |
| solubility | TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C |
| specific activity | ≥800 CDU/mg solid |
| 2-5 FALGPA units/mg solid | |
| storage temp. | −20°C |
| Analysis Note: | Also contains clostripain, nonspecific neutral protease, and tryptic activities. |
| Application: | Clostridiopeptidase A has been used in a study to describe a quantitative method to assess small amounts of collagen based on MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry (MS) subsequent to digestion of collagen. |
| Application: | This product has been used in the isolation of pancreatic ductal adenocarcinoma (PDAC) cell lines and the tissue culture of pancreatic fibroblasts. |
| 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: | Effective release of cells from tissue requires the action of collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca2+) per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. The pH optimum is 6.3-8.8. The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Collagenase treatment can cause some cells to die. Typically, concentrations varying from 0.1 to 5 mg/mL are used for digestion. The duration of the reaction varies from 15 minutes to several hours to yield a satisfactory efficiency of cell dissociation without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preffered and Zn2+ is required for activity. |
| 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: | A stock solution may be prepared by dissolving 0.05-0.1 mg/mL of collagenase in 50 mM TES buffer containing 0.36 mM calcium chloride (TESCA), 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: ≥800 CDU/mg solid; specific activity: 2-5 FALGPA units/mg solid |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 3.4.24.3 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |