Collagenase from Clostridium histolyticum
SIGMA/C8051 - Sigma Blend Type H, ≥1.0 FALGPA units/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.
| caseinase activity | ≥10 units/mg solid |
| form | powder |
| mol wt | 68-130 kDa |
| Quality Level | 200  |
| specific activity | ≥1.0 FALGPA units/mg solid |
| storage temp. | −20°C |
| Application: | Collagenase from Clostridium histolyticum has been used: • to harvest osteosarcoma 143B cells from collagen scaffold • in tissue digestion of islets and carotid artery • in the digestion of chorionic villi • in enzymatic dissociation for ATP-based tumour chemosensitivity assay (ATP-TCA) in solid tumors |
| Biochem/physiol Actions: | Clostridium histolyticum produces two classes of collagenases. Clostridial collagenases are found to be highly effective than mammalian collagenases. It is capable of cleaving at multiple cleavage sites within the collagen triple helix. It is implicated in the bacterial invasion in gas gangrene. |
| 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 both 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 is used for digestion. The duration of reaction varyies 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. This enzyme is tested for the release of hepatocytes at approximately 1 mg/mL conc., in a total volume of 100 mL for each rat liver. |
| 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 |
| 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 | caseinase activity: ≥10 units/mg solid; specific activity: ≥1.0 FALGPA units/mg solid |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 3.4.24.3 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |