LIBTH-RO

Roche

Liberase TH Research Grade

high Thermolysin concentration

Quality Level

form

lyophilized

packaging

pkg of 10 mg (05401135001 [2 x 5 mg])
pkg of 100 mg (05401151001 [2 x 50 mg])

manufacturer/tradename

Roche

parameter

35-37 °C optimum reaction temp.

optimum pH

7.4

shipped in

dry ice

storage temp.

−20°C

General description

Liberase TH Research Grade contains highly purified Collagenase I and Collagenase II. These two collagenase isoforms are blended in a precise ratio to each other and with a high concentration of Thermolysin, a non-clostridial neutral protease.

Application

Liberase TH (Thermolysin High) Research Grade is used for the dissociation of a broad range of tissue types, where high purity of the enzyme blend is necessary for high cell yield and viability. Bacterial by-products, such as endotoxins, are reduced up to several thousand-fold.

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.
  • Increase safety
with an enzyme that is free of mammalian or avian tissue-derived raw materials.

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 Wünsch 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 (Wünsch units/mg, [as determined above]), to obtain Wünsch units/ml. To determine how much Liberase Research Grade Enzyme Blend to use, first multiply your collagenase working concentration (in Wünsch units/ml) times the total volume of your working enzyme solution to obtain the total collagenase activity needed (Wünsch 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. For further information on product stability please visit the Roche Liberase Enzyme website at www.collagenase.com.

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 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 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 Wünsch (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)

Other Notes

For life science research only. Not for use in diagnostic procedures.

Legal Information

Liberase is a trademark of Roche

RIDADR

NONH for all modes of transport

Paul W Burridge et al.
Current protocols in human genetics, 87, 21-21 (2015-10-07)
Since the first discovery that human pluripotent stem cells (hPS cells) can differentiate to cardiomyocytes, efforts have been made to optimize the conditions under which this process occurs. One of the most effective methodologies to optimize this process is reductionist...
K Maiellaro-Rafferty et al.
Journal of molecular and cellular cardiology, 60, 151-160 (2013-05-02)
Nebulette (NEBL) is a sarcomeric Z-disk protein involved in mechanosensing and force generation via its interaction with actin and tropomyosin-troponin complex. Genetic abnormalities in NEBL lead to dilated cardiomyopathy (DCM) in humans and animal models. The objectives of this study...
Francesca Incardona et al.
eLife, 4, e06938-e06938 (2015-06-19)
The Reproducibility Project: Cancer Biology seeks to address growing concerns about reproducibility in scientific research by replicating selected results from a substantial number of high-profile papers in the field of cancer biology published between 2010 and 2012. This Registered report...
Artículos
Enzyme Explorer Key Resource: Collagenase Guide.Collagenases, enzymes that break down the native collagen that holds animal tissues together, are made by a variety of microorganisms and by many different animal cells.
Más información
Protocolos
Your current tissue dissociation protocol, and the extent to which you have been successful with traditional collagenase are good guides for selecting the appropriate Liberase Enzyme Blend. Select the Liberase Enzyme Blend most similar to your tissue dissociation application, or based upon a specific manufacturer and collagenase type.
Más información
Contenido relacionado
Collagenase Guide.Collagenases, enzymes that break down the native collagen that holds animal tissues together, are made by a variety of microorganisms and by many different animal cells.
Más información

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