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
MoreLuciferaseKits
| Catalognumber | Product | Description |
|---|---|---|
| 30085 | FireflyLuciferaseAssayKit2.0 | Flash-typefireflyluciferaseassaykit. |
| 30075 | FireflyLuciferaseAssayKit(Lyophilized) | Fireflyluciferaseassaykitwithlyophilizedassaybufferforeconomicalroomtemperatureshippingandconvenientstorageat-20°C. |
| 99923 | 5XFireflyLysisBuffer | ExtralysisbufferforusewithFireflyLuciferaseAssayKits,onlyrequiredifusingwellsizeslargerthan24-well. |
| 30028 | Steady-Luc™FireflyHTSAssayKit | Glow-typefireflyluciferaseassaywithsignalhalf-lifeofabout3hoursforhigh-throughputscreeninginmulti-wellplates. |
| 30028L | Steady-LucFireflyHTSAssayKit(Lyophilized) | Steady-Luckitwithlyophilizedassaybufferforeconomicalroomtemperatureshippingandconvenientstorageat-20°C. |
| 30082 | RenillaLuciferaseAssayKit2.0 | Flash-typeRenillaluciferaseassaykit. |
| 99912 | 5XPassiveLysisBuffer | ExtralysisbufferforusewithRenillaLuciferaseAssayKits,onlyrequiredifusingwellsizeslargerthan24-well. |
| 30081 | FireflyandRenillaLuciferaseSingleTubeAssayKit | AssaykitformeasuringfireflyandRenillaactivitiessequentiallyinthesamesampleinasingletube. |
| 99821-100mL | 1XPassiveLysisBuffer2.0 | ExtralysisbufferforusewithFirefly&RenillaSingleTubeAssayKits,onlyrequiredifusingwellsizeslargerthan24-well. |
| 30020 | ATP-Glo™BioluminometricCellViabilityAssayKit | Flash-typeluminescence-basedassayformeasuringcellviabilityorATPconcentration. |
| 22003 | MiniCellScrapers | Mini-cellscrapers,usefulforlysingcellsin96-well,48-well,or24-wellplates. |
References
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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.
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8.Matthews,J.C.,Hori,K.andCormier,M.J.(1977)Biochemistry16,85–91.
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