- Производитель:
- Roche
Liberase Purified Enzyme Blends
Liberase enzymes are blends of highly purified enzymes, designed to increase the quality and reproducibility of tissue dissociation, and improve the viability and functionality of isolated cells. Liberase Purified Enzyme Blends replace traditional collagenase, which is a crude and variable fermentation by-product of Clostridium histolyticum.
Liberase Enzyme Technology
Liberase enzyme technology comprises methods for purifying clostridial collagenase isoforms to high specific activity, and for blending them together with high-specific-activity neutral protease in optimal ratios for effective dissociation of primary tissues and cultured cells.
For additional information, such as manufacturing, technology, or storage, see the Liberase Research Grade Insightsdocument. Further reading on the topic can also be found in the Documents tab.
Roche provides special-quality preparations of Liberase Purified Enzyme Blends, manufactured to meet GMP requirements. For more information on the GMP-grade preparations.
Important Notes:
- This product is intended for life science research and in vitro use only. These products are not to be used for diagnostic or clinical applications, such as human islet transplantation.
- All 1st generation Liberase and Liberase Blendzyme products (Liberase HI, CI, RI, PI, Liberase Blendzyme 1, 2, 3, 4) have been replaced by the 2nd generation Liberase Research Grade and Liberase GMP grade enzyme blend portfolios. For instructions concerning the transition from previously used Liberase Enzymes.
This product has been used for the isolation of pancreatic islets from mice.
- Maximize viability and yield of isolated cells with an enzyme blend that has less clostripain and trypsin activity, as well as reduced endotoxin content.
- Count on higher specific activity of the enzyme blend as a result of higher Collagenase I + II purit(determined by HPLC analysis)
- Obtain higher experimental reproducibility due to higher lot-to-lot consistency.
- Increase safety with an enzyme that is free of any mammalian or avian tissue-derived raw materials.
Working concentration: Liberase Research Grade Enzyme Working Concentration
Liberase enzymes have significantly higher specific activities than traditional collagenases. This means that the working concentration of Liberase Research Grade Purified Enzymes, expressed in mg/ml, will be lower than that of traditional collagenase.
When the application is on the Roche list of applications at www.collagenase.com, use the Liberase Research Grade concentration recommended for that application.
When the application is not included on this list, first use Liberase TM Research Grade at a concentration of 0.08–0.28 Wnsch units/ml.
The goal is to determine the best starting concentration of Liberase Research Grade Enzyme Blends. This is a starting point, and the final concentration may vary due to differences in procedure and lot-to-lot differences in traditional collagenase.
Collagenase Working Concentration
Multiply your previous collagenase working concentration (mg/ml) by its specific activity (Wnsch units/mg, as determined above), to obtain Wnsch units/ml. To determine how much Liberase Research Grade Enzyme Blend to use, first multiply your collagenase working concentration (in Wnsch units/ml) times the total volume of your working enzyme solution to obtain the total collagenase activity needed (Wnsch units). Divide the total collagenase activity required by the Liberase Research Grade stock concentration ( “Reconstitution and Storage”). This indicates how many milliliters of Liberase Research Grade Enzyme Blend stock solution to use in your working enzyme solution.
Storage conditions (working solution): Store unused stock solution in single-use aliquots at -15 to -25 °C. Note: Avoid repeated freezing and thawing!
Reconstitute the entire vial. Do not weigh individual aliquots of the lyophilizate. The introduction of moisture into the vial results in a decline in enzymatic activity.
Place vial on ice to rehydrate the lyophilized enzyme.
Gently agitate the vial at 2 to 8 °C until enzyme is completely dissolved (max. 30 min).
Depending on the type of tissue-dissociation buffer used to dissolve Liberase Research Grade Purified Enzyme Blends, slight precipitations may be observed which readily dissolve in the diluted working solution and have no influence on enzyme activity.
Remove an aliquot of the stock solution to prepare the working solution
Reconstitution volume
2 ml (1 vial with 5 mg–10 mg pack size), 10 ml (1 vial with 50 mg–100 mg pack size)
Collagenase Wnsch (units/ml)
13 (1 vial with 5 mg–10 mg pack size), 26 (1 vial with 50 mg–100 mg pack size)
Total Collagenase concentration mg/ml
2.5 (1 vial with 5 mg–10 mg pack size), 5.0 (1 vial with 50 mg–100 mg pack size)
Quality Level | 100 |
form | lyophilized |
packaging | pkg of 10 mg (2x5 mg) |
mfr. no. | Roche |
parameter | 35-37 °C optimum reaction temp. |
optimum pH | 7.4 |
shipped in | dry ice |
pictograms |
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