- Производитель:
- Roche
Кат. номер |
5466202001 |
5401160001 |
Features and Benefits
- 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 purity (determined by HPLC analysis).
- Obtain higher experimental reproducibility due to higher lot-to-lot consistency.
Preparation Note
Stabilizers: Calcium
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!
Reconstitution
Reconstitute the lyophilized enzyme with injection-quality sterile water or tissue-dissociation buffer. Do not add serum or other components that may influence enzyme activity, such as albumin or protease inhibitors, to the dissociation buffer. Enzyme stability is reduced at higher concentrations and warmer temperatures (4 °C). Avoid both the above conditions.
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 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)
Liberase is a trademark of Roche
Roche is a registered trademark of Roche Molecular Systems, Inc.
Quality Level | 100, |
form | lyophilized |
packaging | pkg of 100 mg (05466202001 [2 x 50 mg]), pkg of 10 mg (05401160001 [2 x 5 mg]) |
Торговая марка | Roche |
parameter | 35-37 °C optimum reaction temp. |
optimum pH | 7.4 |
shipped in | dry ice |
storage temp. | 20°C |
RIDADR | NONH for all modes of transport |
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