05401020001

Roche

Liberase TL Research Grade

low Thermolysin concentration

Quality Level

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

General description

The most commonly used method for decellularization procedures, such as tissue dissociation and cell harvesting, is based on the use of proteolytic enzymes. Proteases disrupt the extracellular matrix of tissue to allow the release of individual cells or the harvesting of cultured cells for transfer. The goal of cell isolation is to maximize the yield of dissociated cells that are viable and functionally active.

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1st generation Liberase and Liberase Blendzyme products (Liberase HI, CI, RI, PI, Liberase Blendzyme 1, 2, 3, 4) have been replaced by the 2nd generation Liberase Research Grade and Liberase GMP grade enzyme blend portfolios.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.

Application

Liberase TL (Thermolysin Low) 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.
This product has been used for the isolation of pancreatic islets from mice.

Features and Benefits

Liberase TL 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 low concentration of Thermolysin, a non-clostridial neutral protease.
  • 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.

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 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 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)

Storage and Stability

Store at -15–-25 °C. (Store dry. Please note that the expiration date is not printed on the individual vials.)

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

Signal Word

Danger

Hazard Statements

RIDADR

NONH for all modes of transport

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Christine M Kusminski et al.
Diabetes, 65(6), 1534-1555 (2016-02-21)
Mitochondrial metabolism plays an integral role in glucose-stimulated insulin secretion (GSIS) in β-cells. In addition, the diabetogenic role of glucagon released from α-cells plays a major role in the etiology of both type 1 and type 2 diabetes because unopposed...
Erik J Zmuda et al.
Journal of visualized experiments : JoVE, (50)(50), doi:10-doi:10 (2011-04-29)
Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize their blood glucose levels, islet transplantation has been proposed to be a potential treatment for type 1 diabetes. More...
Risheng Ye et al.
eLife, 3, doi:10-doi:10 (2014-10-24)
As an adipokine in circulation, adiponectin has been extensively studied for its beneficial metabolic effects. While many important functions have been attributed to adiponectin under high-fat diet conditions, little is known about its essential role under regular chow. Employing a...
Kavaljit H Chhabra et al.
American journal of physiology. Endocrinology and metabolism, 304(8), E874-E884 (2013-03-07)
An overactive renin-angiotensin system (RAS) is known to contribute to type 2 diabetes mellitus (T2DM). Although ACE2 overexpression has been shown to be protective against the overactive RAS, a role for pancreatic ACE2, particularly in the islets of Langerhans, in...
Articles
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.
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Protocols
Liberase™ TL Research Grade Protocol & Troubleshooting
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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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