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DescriptionFireflyluciferaseiswidelyusedasareporterforstudyinggeneregulationandfunction,andforpharmaceuticalscreening1,2.Itisaverysensitivegeneticreporterduetothelackofanyendogenousactivityinmammaliancellsortissues3,4.TheFireflyluciferaseisa62,000Daltonprotein,whichisactiveasamonomeranddoesnotrequiresubsequentprocessingforitsactivity.TheenzymecatalyzesATP-dependentD-luciferinoxidationbyoxygenintooxyluciferinwithemissionoflightcenteredon560nm(Figure1).Aswithmanyenzymes,FireflyluciferasefollowsMichaelis-Mentenkineticsand,asaresult,maximumlightoutputisnotachieveduntilthesubstrateandco-factorsarepresentinlargeexcess.Whenassayedundertheseconditions,lightemittedfromthereactionisdirectlyproportionaltothenumberofluciferaseenzymemolecules.ThisFireflyluciferaseassaykitisdesignedfordetectionandquantificationoftheFireflyluciferasereporterenzymefromculturedcellsinasimple,efficient,andlinearfashion(Figure3).Figure1.BioluminescentreactioncatalyzedbyFireflyluciferase.RenillaLuciferasehasbeenusedasareporterforstudyinggeneregulationandfunctioninvitroandinvivo5,6.Recently,RenillaLuciferasehasbeenwidelyusedinmultiplextranscriptionalreporterassaysorasanormalizingtransfectioncontrolforFireflyluciferaseassay6,7.Renillaluciferase,amonomeric36,000Daltonprotein,catalyzescoelenterazineoxidationbyoxygent

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