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.