Collagenase D
ROCHE/COLLD-RO - from Clostridium histolyticum
Product Type: Chemical
Collagenase D
| application(s) | cell analysis |
| biological source | Clostridium histolyticum |
| collagenase activity | >0.15 U/mg lyophilizate |
| color | light brown |
| concentration | (working concentration: 0.5 - 2.5 mg/ml) |
| foreign activity | Clostripain 0.060 U/mg |
| Proteases 8 U/mg ( Azocoll) | |
| Trypsin <0.2 U/mg (with BAEE) | |
| form | lyophilized |
| manufacturer/tradename | Roche |
| NCBI accession no. | WP_171012037   |
| optimum pH | 6.0-8.0 |
| packaging | pkg of 100 mg (11088858001) |
| pkg of 2.5 g (11088882001) | |
| pkg of 500 mg (11088866001) | |
| Quality Level | 100 |
| solubility | water: soluble |
| sterility | non-sterile |
| storage temp. | 2-8°C |
| technique(s) | tissue processing: suitable |
| Application: | Collagenase from C. histolyticum is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single cell suspensions for the establishment of primary cell culture systems. Collagenase D is recommended when functionality and integrity of cell-surface proteins are important. Clostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically. |
| Biochem/physiol Actions: | Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well. |
| Features and Benefits: | Contents Lyophilizate, nonsterile |
| General description: | Collagenase D is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization. |
| Other Notes: | Collagenase from Roche is assayed in Wünsch units (1 μmol of product formed per minute at +25 °C with Wünsch substrate). Frequently, collagenase activities are given in Mandl units (1 μmol leucine liberated from collagen in 5 hours at +37 °C). Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wünsch U/mg and 250 Mandl U/mg). |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Physical form: | Lyophilizate, nonsterile. |
| Preparation Note: | Activator: Ca2+ Working concentration: 0.5 - 2.5mg/ml Storage conditions (working solution): -15 to -25°C Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25°C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution. Inhibitors: Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol Collagenase is not inhibited by serum. Clostripain inhibitors: TLCK Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean) Enzyme activity: Collagenase activity: >0.15 U/mg (according to Wünsch) (+25°C, 4-phenyl-azobenzyl-oxycar Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity Collagenase D has a normal to high collagenase activity and very low tryptic activity (usually <0.2 units/mg, BAEE as substrate). |
| Preparation Note: | Keep protected from light |
| Preparation Note: | Reconstitution in any balanced salt solution (e.g., HBSS) |
| Symbol |   GHS07,GHS08 |
| Signal word | Danger |
| Hazard statements | H315 - H319 - H334 - H335 |
| Precautionary statements | P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| activity | collagenase activity: >0.15 U/mg lyophilizate |
| Storage Temp. | 2-8°C |
| Enzyme Commission (EC) Number | 3.4.24.3 ( BRENDA | IUBMB ) |
| UNSPSC | 12352204 |