Collagenase from Clostridium histolyticum
SIGMA/C9891 - Type IA, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid, For general use
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 |
| form | essentially salt-free, lyophilized powder |
| mol wt | 68-130 kDa |
| Quality Level | 200  |
| shipped in | wet ice |
| solubility | TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble |
| specific activity | ≥125 CDU/mg solid |
| 0.5-5.0 FALGPA units/mg solid | |
| storage temp. | −20°C |
| technique(s) | cell culture | mammalian: suitable |
| single cell analysis: suitable |
| Application: | The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, and lung epithelial tissues. It is also useful in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocytes. The enzyme from Sigma has been used for the digestion of type I collagen from bovine trabecular and cortical bones. It has also been used as a positive control in FALGPA assay on Group B Streptococci cells. Suitable for use in preparation of single cell suspension for sequencing. |
| 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 ranges from 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 reaction varies from 15 minutes to several hours for 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. The enzyme recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid. Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraace |
| 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 product is equivalent to first 40% ammonium sulfate fraction of Mandl, I., et al., J. Clin. Invest., 32, 1323 (1953). 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: ≥125 CDU/mg solid; specific activity: 0.5-5.0 FALGPA units/mg solid |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 3.4.24.3 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |