I.U.B.:3.4.24.3

CollagenaseRawMaterialCertification 

Crudecollagenasepreparationscontainseveralisoformsoftwodifferentcollagenases,asulfhydrylprotease,clostripain,atrypsin-likeenzyme,andanaminopeptidase.Thiscombinationofcollagenolyticandproteolyticactivitiesiseffectiveatbreakingdownintercellularmatrices,theessentialpartoftissuedissociation.OnecomponentofthecomplexisahydrolyticenzymewhichdegradesthehelicalregionsinnativecollagenpreferentiallyattheY-GlybondinthesequencePro-Y-Gly-Pro,whereYismostfrequentlyaneutralaminoacid.ThiscleavageyieldsproductssusceptIBLetofurtherpeptidasedigestion.Crudecollagenaseisinhibitedbymetalchelatingagentssuchascysteine,EDTAoro-phenanthrolinebutnotDFP.Itisalsoinhibitedbyα2-macroglobulin,alargeplasmaglycoprotein.Ca²⁺isrequiredforenzymeactivity.Particularenzymaticprofilesofeachcollagenasehavebeencorrelatedwiththetissuesfromwhichthecellsforstudywereobtained(orwiththeusestowhichthecellsareput)andasaresultofthecorrelationsseveraltypesofcrudecollagenaseshavebeenestablishedbyWorthington:Types1,2,3,and4.

•Type1crudecollagenasehastheoriginalbalanceofcollagenase,caseinase,clostripainandtrypticactivities.
•Type2containshigherrelativelevelsofproteaseactivity,particularlyclostripain.
•Type3containslowestlevelsofsecondaryproteases.
•Type4isdesignedtobeespeciallylowintrypticactivitytolimitdamagetomembraneproteinsandreceptors.
•Purifiedcollagenase,Code:CLSPA,containsminimalsecondaryproteolyticactivitiesalongwithhighcollagenaseactivity.

Worthingtonalsooffers0.22µmfilteredpreparationsofeachtypeinpre-packagedformfordirectreconstitutionanduseincellisolationandcultureprocedures.Correlationsbetweenenzymetypeandeffectivenesswithdifferenttissueshavebeengood,butnotperfect,dueinparttothevariableparametersofuse.Neverthelessmostresearchersconsiderthetissue-typingofcrudecollagenaselotstobeavaluableservice.AdetaileddescriptionoftheWorthingtoncollagenaseandcontaminantassayscanbefoundintheWorthingtonEnzymeManual.Inaddition,tissuespecificreferencesanddetailedisolationconditionscanbefoundintheWorthingtonTissueDissociationGuide.

ThecollagenaseassayisamodificationofMandlwhereincollagenaseisincubatedforfivehourswithnativecollagenandtheextentofcollagenbreakdownisdeterminedusingtheMooreandStein,JBC,176,367,(1948)colorimetricninhydrinmethod.Aminoacidsreleasedareexpressedasmicromolesleucinepermilligramcollagenase.

Uses:Crudecollagenasesarewidelyusedinenzymaticprimarycellisolationandtissuedissociationprocedures.MostresearchersemployeithercrudecollagenasepreparationssuchasTypes1,2,3,and4orchromatographicallypurifiedcollagenase(Code:CLSPA);thelatterusuallycombinedwithsecondaryenzymessuchaselastase,hyaluronidase,etc.Forbestresults,theprecisemixtureofproteolyticactivitiesmustbetailoredtothetissuetobedissociated.Correlationsbetweentypeandeffectivenesswithdifferenttissueshavebeengood,butnotperfect,andmaybedependentpartlyonparametersofuseandobjectivesaswellaslot-to-lotvariations.FormoreinformationseetheCollagenaseSamplingPrograminformation.WorthingtonalsopublishesaTissueDissociationGuidewhichprovidesadditionalinformationregardingtheenzymesusedfortheseapplicationsandspecificreferencesfornumerouscellandtissuetypes.

UnitDefinition:OneUnitreleasesonemicromoleofL-leucineequivalentsfromcollagenin5hoursat37°C,pH7.5

RelatedProducts:

Collagen(CL)
DeoxyribonucleaseI(DP/D/DCLS/D2/DPFF/DPRF)
Elastase(ES/ESL/ESFF)
Hyaluronidase(HSE/HSEP)
NeutralProtease(Dispase,NPRO)
Papain(PAP/PAPL/PAP2)ProteinaseK(PROK)
Trypsin(TL/TRL/TRL3/TRLS/TRTPCK)
TrypsinInhibitors(LBI/OI/SI/SIC)

CellIsolationOptimizingSystem(CIT)
HepatocyteIsolationSystem(HIS)
NeonatalCardiomyocyteIsolationSystem(NCIS)
PapainDissociiationSystem(PDS/PDS2)

Worthington于1947年创办于美国，是一家通过的ISO9001认证的家族企业。 主要生产用于生物技术和生命科学研究，诊断，生物制药和生物加工应用的高质量纯化酶，蛋白质，核酸和试剂盒。我们为所有类型的生命科学应用生产提供优质产品的声誉是70多年经验的结晶。Worthington的内部全方位服务能力包括初始提取，发酵，蛋白质纯化，冻干和包装。所有流程都有完整的文件记录，可从原材料到最终装运进行追溯。

专注领域：

l  酶

l  蛋白质

l  核酸

l  细胞分离试剂盒

l  无动物源性的酶/蛋白质

作为世界上最主要的酶类供应商，以及细胞分离酶的专家级公司，公司的核心酶产品包括：

l  胶原蛋白酶

l  脱氧核糖核酸酶I

l  弹性蛋白酶

l  木瓜蛋白酶