FireflyluciferasehasbeenusedasasensitivereportertostudyawiderangeofBIOLOGicalresponses.HoweverthechangeoftheexpressionofFireflyluciferasereportercanbeduetoaglobaleffectinsteadofaspecificeffect.TheaccuracyofFireflyluciferasereportercanbeimprovedbynormalizingtoacontrolreporter,suchasRenillaluciferasereporter,inthesamesample.
TheDualLuciferase(Firefly-Renilla)AssaySystemisdesignedtobeusedforhigh-throughput,rapidquantitationofbothFireflyandRenillaluciferasesfromasinglesampleinmammaliancellculture.TheFireflyLuciferaseReagentisfirstaddedtothecellsinmediumdirectly.Thisreagentlysesthecellsandcontainsasubstrateforfireflyluciferasetoproducefireflyluciferaseluminescence.Next,theRenillaLuciferaseReagentisaddedtothesamewell.Itquenchesthefireflyluciferaseluminescenceandprovidesthesubstrateforrenillaluciferasetoproducerenillaluciferaseluminescence.Thelightproductionofbothreactionscanbeconvenientlymeasuredonaluminometer.Thisassaysystemhasseveralfeatures:
•Sensitive–highlysensitivedetectionoffireflyluciferaseactivityandRenillaluciferaseactivity.
•Stable–theluciferasesignaloutputisstableformorethanonehour,providingflexibilitywithregardtoincubationtime
•High-throughput–simplehomogeneousprotocolminimizeshandlingstepstosupporthigh-throughputscreeningapplications
•Compatibility–workswellwithavarietyofcommonmediacontaining0-10%serumandphenolred.
COMPONENTS:
•High-throughputdrugscreeningusingFirefly-Renilladualluciferasereporter.
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