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Adipogen/Ubiquitin Conjugation Kit/AG-44T-0132-KI01/1 Kit
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Specifications&Handling:ProductDataSynonymsUBB;UbiquitinBPropertiesCrossreactivityRabbitKitContains200µgofConugationFractionA200µgofConugationFractionB50µlofUbiquitinSolution150µlof10XEnergySolutionNOTE:Thiskitsuppliesenoughmaterialsfor10x50µlreactions.OtherProductDataUse:Thiskitisfortheformationofubiquitinatedsubstrateproteins.Theenzymessuppliedrepresentthefullcomplimentofpurifiedconjugationenzymes(E1,E2s,andE3s)thatarefoundinFractionII.ConjugationFractionAcontainspredominantlyE1andE2enzymes,whileConjugationFractionBcontainspredominantlyE3anddeubiquitinatingenzymes.Theenzymeshavebeentestedandshowntoworkwithtypical[125I]-labeledsubstrateproteinssuchaslysozymeandβ-lactoglobulin.ThisconjugationfractionalsocontainsubiquitinC-terminalhydrolasesandthereforetheadditionofUbiquitinAldehyde(AG-40T-0511)isrecommendedfortheinhibitionofUCHstoimproveoverallconjugateyield.Thesuppliedfractionsdonotcontain20Sor26Sproteasomesoranyotherproteindegradationactivity.IfthesubstratebeingconjugatedrequiresE2orE3enzymesnotfoundinFractionII,thereactioncanbesupplementedwithenzymesfromFractionI(Prod.No.AG-40T-0344).OriginManufacturedbyBostonBiochemProductTypeKitShippingandHandlingShippingDRYICEShortTermStorage-20°CLongTermStorage-80°CHandlingAdviceAliquottoavoidfreeze/thawcycles.Use/StABIlityStableforatleast6monthsafterreceiptwhenstoredat-80°C.DocumentsManualDownloadDescription:Ubiquitinisa76aminoacid(aa)proteinthatisubiquitouslyexpressedinalleukaryoticorganisms.ubiquitinishighlyconservedwith96%aasequenceidentitysharedbetweenhumanandyeastubiquitin,and100%aasequenceidentitysharedbetweenhumanandmouseubiquitin.Inmammals,fourubiquitingenesencodefortwoubiquitin-ribosomalfusionproteinsandtwopoly-ubiquitinproteins.Cleavageoftheubiquitinprecursorsbydeubiquitinatingenzymesgivesrisetoidenticalubiquitinmonomerseachwithapredictedmolecularweightof8.6kDa.ConjugationofubiquitintotargetproteinsinvolvestheformationofanisopeptidebondbetweentheC-terminalglycineresidueofubiquitinandalysineresidueinthetargetprotein.Thisprocessofconjugation,referredtoasubiquitinationorubiquitylation,isamulti-stepprocessthatrequiresthreeenzymes:aubiquitin-activating(E1)enzyme,aubiquitin-conjugating(E2)enzyme,andaubiquitinligase(E3).ubiquitinationisclassicallyrecognizedasamechanismtotargetproteinsfordegradationandasaresult,ubiquitinwasoriginallynamedATP-dependentProteolysisFactor1(APF-1).Inadditiontoproteindegradation,ubiquitinationhasbeenshowntomediateavarietyofBIOLOGicalprocessessuchassignaltransduction,endocytosis,andpost-endocyticsorting.
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