NOVAGEN_Thrombin_kit

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1、ThrombinKitsTableofContentsAbouttheKits.......................................................................................2Description2Components2ThrombinCleavage................................................................................3Smallscaleoptimization3Scale-up4Factorst

2、hataffectthrombinactivity4Monitoringcleavage5BiotinylatedThrombinCapture..................................................................5References...........................................................................................6Copyright1992–1998byNovagen,Inc.Allrightsreser

3、ved.S•Tag™,T7•Tag®,His•Bind®,pBAC™,CBD•Tag™,PerfectProtein™,theNovagenlogoandnamearetrademarksofNovagen,Inc.Orders:800526-7319TechnicalService:800207-0144TB1888/98Novagen1ThrombinKitsAbouttheKitsThrombinCleavageCaptureKit69022-3BiotinylatedThrombin69672-3Thrombin,RestrictionGrade69671-3

4、DescriptionThrombinisanendoproteasethatnaturallyfunctionsasabloodclottingfactortoconvertfibrinogentofibrin.Humanthrombinisoneofthemostactivesite-specificproteases,whichisanadvantagebecauseaverylowmassratioofenzymetotargetproteinisneededforefficientcleavage.Althoughnosingleconsensusseque

5、ncedescribesthespecificityofthrombincutting,cleavagefrequentlyoccursaftertheproline-arginineresiduepairwhenproperlyexposedinathree-dimensionalstructure.Novagen’sBiotinylatedThrombinandRestrictionGradeThrombinarefunctionallytestedforspecificcleavageofthesequenceLeuValProArg¯GlySerinrecom

6、binantfusionproteins.Thisthrombinrecognitionsequenceisencodedbymanypopularexpressionvectors,includingavarietyofNovagen’svectors.Thecleavagesiteisusuallylocatedbetweenthecloningsitesandothervector-encodedfusionpeptidesequences,whichallowsproteolyticremovalofthefusedpeptidefromtheexpresse

7、dtargetprotein.BiotinylatedThrombinissimilartoRestrictionGradeThrombin,butithasbiotincovalentlyattachedforeasyremovaloftheenzymefromcleavagereactionsusingimmobilizedstreptavidin.Boththrombinproductsarerigorouslytestedforspecificcleavageofarecombinantfusionproteinandarefunctiona