10/08/2015

LiberaseTLResearchGradelowThermolysinconcentration|SigmaAldrich

BrazilIncio 05401020001LiberaseTLResearchGrade

05401020001ROCHE

LiberaseTLResearchGrade
lowThermolysinconcentration

Propriedades
form

lyophilized

mfr.no.

Roche

packaging

pkgof10mg(2x5mg)

limit

3537Coptimumreactiontemp.

optimumpH

7.4

Descrio
Application
LiberaseTL(ThermolysinLow)ResearchGradeisusedforthedissociationofabroadrangeoftissuetypes,wherehighpurityoftheenzymeblendis
necessaryforhighcellyieldandviability.Bacterialbyproducts,suchasendotoxins,arereduceduptoseveralthousandfold.
FeaturesandBenefits
LiberaseTLResearchGradecontainshighlypurifiedCollagenaseIandCollagenaseII.Thesetwocollagenaseisoformsareblendedinapreciseratiotoeach
otherandwithalowconcentrationofThermolysin,anonclostridialneutralprotease.
Maximize viability and yield of isolated cells withanenzymeblendthathaslessclostripainandtrypsinactivity,aswellasreducedendotoxin
content.
Count on higher specific activity of the enzyme blendasaresultofhigherCollagenaseI+IIpurit(determinedbyHPLCanalysis)
Obtain higher experimental reproducibilityduetohigherlottolotconsistency.
Increase safetywithanenzymethatisfreeofanymammalianoraviantissuederivedrawmaterials.
Generaldescription
DownloadtheCofA,MSDSandIFUspecifictoyourproductlotandformat

Themostcommonlyusedmethodfordecellularizationprocedures,suchastissuedissociationandcellharvesting,isbasedontheuseofproteolyticenzymes.
Proteasesdisrupttheextracellularmatrixoftissuetoallowthereleaseofindividualcellsortheharvestingofculturedcellsfortransfer.Thegoalofcellisolation
istomaximizetheyieldofdissociatedcellsthatareviableandfunctionallyactive.
Liberase Purified Enzyme Blends
Liberaseenzymesareblendsofhighlypurifiedenzymes,designedtoincreasethequalityandreproducibilityoftissuedissociation,andimprovetheviability
andfunctionalityofisolatedcells.LiberasePurifiedEnzymeBlendsreplacetraditionalcollagenase,whichisacrudeandvariablefermentationbyproductof
Clostridiumhistolyticum.
Liberase Enzyme Technology
Liberaseenzymetechnologycomprisesmethodsforpurifyingclostridialcollagenaseisoformstohighspecificactivity,andforblendingthemtogetherwith
highspecificactivityneutralproteaseinoptimalratiosforeffectivedissociationofprimarytissuesandculturedcells.
Foradditionalinformation,suchasmanufacturing,technology,orstorage,seetheLiberaseResearchGradeInsightsdocument.Furtherreadingonthetopic
canalsobefoundintheDocumentstab.
RocheprovidesspecialqualitypreparationsofLiberasePurifiedEnzymeBlends,manufacturedtomeetGMPrequirements.FormoreinformationontheGMP
gradepreparations.
Important Notes:
Thisproductisintendedforlifescienceresearchandinvitrouseonly.Theseproductsarenottobeusedfordiagnosticorclinicalapplications,suchas
humanislettransplantation.
All1stgenerationLiberaseandLiberaseBlendzymeproducts(LiberaseHI,CI,RI,PI,LiberaseBlendzyme1,2,3,4)havebeenreplacedbythe2nd
generationLiberaseResearchGradeandLiberaseGMPgradeenzymeblendportfolios.Forinstructionsconcerningthetransitionfrompreviouslyused
LiberaseEnzymes.
OtherNotes
Forlifescienceresearchonly.Notforuseindiagnosticprocedures.
PreparationNote
Stabilizers: Calcium
Working concentration: Liberase Research Grade Enzyme Working Concentration

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Liberaseenzymeshavesignificantlyhigherspecificactivitiesthantraditionalcollagenases.ThismeansthattheworkingconcentrationofLiberaseResearch
GradePurifiedEnzymes,expressedinmg/ml,willbelowerthanthatoftraditionalcollagenase.
WhentheapplicationisontheRochelistofapplicationsatwww.collagenase.com,usetheLiberaseResearchGradeconcentrationrecommendedforthat
application.
Whentheapplicationisnotincludedonthislist,firstuseLiberaseTMResearchGradeataconcentrationof0.080.28Wnschunits/ml.
ThegoalistodeterminethebeststartingconcentrationofLiberaseResearchGradeEnzymeBlends.Thisisastartingpoint,andthefinalconcentrationmay
varyduetodifferencesinprocedureandlottolotdifferencesintraditionalcollagenase.
Collagenase Working Concentration
Multiplyyourpreviouscollagenaseworkingconcentration(mg/ml)byitsspecificactivity(Wnschunits/mg,[asdeterminedabove]),toobtainWnsch
units/ml.TodeterminehowmuchLiberaseResearchGradeEnzymeBlendtouse,firstmultiplyyourcollagenaseworkingconcentration(inWnschunits/ml)
timesthetotalvolumeofyourworkingenzymesolutiontoobtainthetotalcollagenaseactivityneeded(Wnschunits).Dividethetotalcollagenaseactivity
requiredbytheLiberaseResearchGradestockconcentration(ReconstitutionandStorage
).ThisindicateshowmanymillilitersofLiberaseResearchGrade
EnzymeBlendstocksolutiontouseinyourworkingenzymesolution.

Storage conditions (working solution): Storeunusedstocksolutioninsingleusealiquotsat15to25C.Forfurtherinformationonproductstability,

pleasevisittheRocheLiberaseEnzymewebsiteatwww.collagenase.com.
Note:Avoidrepeatedfreezingandthawing!
Reconstitution
Reconstitutethelyophilizedenzymewithinjectionqualitysterilewaterortissuedissociationbuffer.Donotaddserumorothercomponentsthatmayinfluence
enzymeactivity,suchasalbuminorproteaseinhibitors,tothedissociationbuffer.Enzymestabilityisreducedathigherconcentrationsandwarmer
temperatures(4C).Avoidboththeaboveconditions.
Reconstitutetheentirevial.Donotweighindividualaliquotsofthelyophilizate.Theintroductionofmoistureintothevialresultsinadeclineinenzymatic
activity.
Placevialonicetorehydratethelyophilizedenzyme.
Gentlyagitatethevialat2to8Cuntilenzymeiscompletelydissolved(max.30min).
DependingonthetypeoftissuedissociationbufferusedtodissolveLiberaseResearchGradePurifiedEnzymeBlends,slightprecipitationsmaybeobserved
whichreadilydissolveinthedilutedworkingsolutionandhavenoinfluenceonenzymeactivity.
Removeanaliquotofthestocksolutiontopreparetheworkingsolution
Reconstitution volume
2ml(1vialwith5mg10mgpacksize),10ml(1vialwith50mg100mgpacksize)
Collagenase Wnsch (units/ml)
13(1vialwith5mg10mgpacksize),26(1vialwith50mg100mgpacksize)
Total Collagenase concentration [mg/ml]
2.5(1vialwith5mg10mgpacksize),5.0(1vialwith50mg100mgpacksize)
LegalInformation
LiberaseisatrademarkofRoche

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