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LIBTH-RO

Roche

Liberase TH Research Grade

lyophilized, suitable for tissue processing, optimum pH 7.4

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biological source

Bacillus polymyxa (dispase)
Bacillus sp. (thermolysin - Bacillus thermoproteolyticus)
Clostridium histolyticum (collagenase)

Quality Level

description

high Thermolysin concentration

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.

technique(s)

tissue processing: suitable

optimum pH

7.4

storage temp.

−20°C

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This Item
LIBTM-ROLIBDH-ROCOLLH-RO
Liberase™ TH Research Grade lyophilized, suitable for tissue processing, optimum pH 7.4

LIBTH-RO

Liberase TH Research Grade

Liberase™ TM Research Grade medium Thermolysin concentration

LIBTM-RO

Liberase TM Research Grade

Liberase™ DH Research Grade lyophilized, optimum pH 7.4, optimum reaction temp. 35-37 °C

LIBDH-RO

Liberase DH Research Grade

Collagenase H from Clostridium histolyticum

COLLH-RO

Collagenase H

technique(s)

tissue processing: suitable

technique(s)

single cell analysis: suitable, tissue processing: suitable

technique(s)

activity assay: suitable

technique(s)

tissue processing: suitable

form

lyophilized

form

lyophilized

form

lyophilized

form

lyophilized

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

2-8°C

optimum pH

7.4

optimum pH

7.4

optimum pH

7.4

optimum pH

6.0-8.0

Quality Level

100

Quality Level

100

Quality Level

100

Quality Level

100

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

pictograms

Exclamation markHealth hazard

signalword

Danger

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

target_organs

Respiratory system

Storage Class

11 - Combustible Solids

wgk_germany

WGK 1

flash_point_f

does not flash

flash_point_c

does not flash


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Protocols

Liberase Enzyme Blend selection depends on tissue dissociation protocol and success with collagenase.

Related Content

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.

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