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Biotium/Firefly & Renilla Luciferase Single Tube Assay Kit/30081-1/150-assays

ThiskitisdesignedtomeasureFireflyandRenillaluciferaseactivitiesintransfectedcellreporterassays.Fireflyluciferaseiswidelyusedasareporterforstudyinggeneregulationandfunction,andforpharmaceuticalscreening.

ThiskitisdesignedtomeasureFireflyandRenillaluciferaseactivitiesintransfectedcellreporterassays.Fireflyluciferaseiswidelyusedasareporterforstudyinggeneregulationandfunction,andforpharmaceuticalscreening.Itisaverysensitivegeneticreporterduetotheabsenceofendogenousluciferaseactivityinmammaliancellsortissues.Renillaluciferasehasbeenusedasareportergeneforstudyinggeneregulationandfunctioninvitroandinvivo. Itcommonlyisusedinmultiplextranscriptionalreporterassaysorasanormalizingtransfectioncontrolforfireflyluciferaseassays.

TheFirefly&RenillaLuciferaseSingleTubeAssayisacombinedluciferaseassayallowingsequentialmeasurementofFireflyandRenillaluciferaseactivityinthesamesamplewithhighsensitivityandlinearity(Figure1).Fireflyluciferaseactivityismeasuredfirst,thenRenillaLuciferaseAssayBuffer2.0isaddedtothesampletosimultaneouslyquenchfireflyluciferaseactivityandmeasureRenillaluciferaseactivity.TheRenillaLuciferaseAssayBuffer2.0quenchesthefireflyluciferaseactivitytothelevelofuntransfectedcells,allowingsequentialmeasurementoffireflyandRenillaluciferaseactivityinthesamesample(Figure2).Thisisaflash-typeassaythatrequiresluminescencetobemeasuredimmediatelyafteraddingluciferasedetectionreagentstoyoursample.

ThiskitisanewandimprovedreplacementfortheFirefly&RenillaDualLuciferaseAssayKit(catalogno.30005).Inadditiontoprovidingasimplified,singletubeassay,thekitfeaturesBiotium’sAquaphile™coelenterazine,awatersolublesubstratethatcanbestoredat-20°Cwithminimalevaporation,unlikemethanolsolutionsofcoelenterazine.

TheFirefly&RenillaLuciferaseSingleTubeAssayfeaturesBiotium’swater-solubleAquaphile™coelenterazine.Italsoincludesournew1XPassiveLysisBuffer2.0,whichisready-to-usewithoutdilutionforcelllysis.ThebufferalsocanbeusedtodiluterecombinantfireflyorRenillaluciferaseenzymeswithouttheneedtoaddBSAtostABIlizetheenzymes.

Thestatednumberofassaysisbasedon96-wellplateformat.Thekitsincludeenough1XPassiveLysisBuffer2.0toperformthestatednumberofassaysinplatesizesrangingfrom96-wellto24-well.Ifneeded,additional1XPassiveLysisBuffer2.0 (catalogno.99821)canbepurchasedseparately.


Kitcomponents:

  • 1XPassiveLysisBuffer2.0
  • FireflyLuciferaseAssayBuffer2.0
  • D-Luciferin
  • RenillaLuciferaseAssayBuffer2.0
  • AquaphileCoelenterazine

Features:

  • Highsensitivityandlinearity
  • Convenientsingletubeformat
  • Watersolublesubstrates

Assay schematic

 

 

ExampleofFirefly&RenillaLuciferaseSingleTubeAssayusinglysatesfromuntransfectedHeLacellsorcellstransfectedwitheitherfireflyluciferasealone(FirelfyOnly)orco-transfectedwithfireflyandRenillaluciferases(Firefly+Renilla).Incellstransfectedwithfireflyonly,theRenillasignalistheresidualfireflyluminescenceafteraddingRenillaworkingsolutiontothereaction.LuminescencewasmeasuredonaPromegaGlomax®20/20singletubeluminometerusingtheDualGlo®programwithintegrationtimeof1second.

MoreLuciferaseKits

CatalognumberProductDescription
30085FireflyLuciferaseAssayKit2.0Flash-typefireflyluciferaseassaykit.
30075FireflyLuciferaseAssayKit(Lyophilized)Fireflyluciferaseassaykitwithlyophilizedassaybufferforeconomicalroomtemperatureshippingandconvenientstorageat-20°C.
999235XFireflyLysisBufferExtralysisbufferforusewithFireflyLuciferaseAssayKits,onlyrequiredifusingwellsizeslargerthan24-well.
30028Steady-Luc™FireflyHTSAssayKitGlow-typefireflyluciferaseassaywithsignalhalf-lifeofabout3hoursforhigh-throughputscreeninginmulti-wellplates.
30028LSteady-LucFireflyHTSAssayKit(Lyophilized)Steady-Luckitwithlyophilizedassaybufferforeconomicalroomtemperatureshippingandconvenientstorageat-20°C.
30082RenillaLuciferaseAssayKit2.0Flash-typeRenillaluciferaseassaykit.
999125XPassiveLysisBufferExtralysisbufferforusewithRenillaLuciferaseAssayKits,onlyrequiredifusingwellsizeslargerthan24-well.
30081FireflyandRenillaLuciferaseSingleTubeAssayKitAssaykitformeasuringfireflyandRenillaactivitiessequentiallyinthesamesampleinasingletube.
99821-100mL1XPassiveLysisBuffer2.0ExtralysisbufferforusewithFirefly&RenillaSingleTubeAssayKits,onlyrequiredifusingwellsizeslargerthan24-well.
30020ATP-Glo™BioluminometricCellViabilityAssayKitFlash-typeluminescence-basedassayformeasuringcellviabilityorATPconcentration.
22003MiniCellScrapersMini-cellscrapers,usefulforlysingcellsin96-well,48-well,or24-wellplates.

References

1.Alam,J.andJ.L.Cook.1990.Anal.Biochem.188:245-254.
2.Bronstein,I.,etal.1994.Anal.Biochem.219:169-181.
3.Gould,S.J.andS.Subramani.1988.Anal.Biochem.175:5-13.
4.Brasier,A.R.,etal.1989.BioTechniques.7:1116-1122.
5.BhaumikS.etal.(2004)JBiomedOpt.9,578-86.
6.MatijasevicZ.etal.(2001)Carcinogenesis.22,661-4.
7.NieuwenhuijsenBW.etal.(2004)JBiomolScreen.8,676-84.
8.Matthews,J.C.,Hori,K.andCormier,M.J.(1977)Biochemistry16,85–91.

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