Fireflyluciferaseiswidelyusedasareporterforstudyinggeneregulationandfunction,andforpharmaceuticalscreening.Itisaverysensitivegeneticreporterduetotheabsenceofendogenousluciferaseactivityinmammaliancellsortissues.Fireflyluciferaseisa62,000Daltonprotein,whichisactiveasamonomeranddoesnotrequiresubsequentprocessingforitsactivity.TheenzymecatalyzesATP-dependentD-luciferinoxidationtooxyluciferin,producinglightemissioncenteredat560nm.FireflyluciferasefollowsMichaelis-Mentenkineticsand,asaresult,maximumlightoutputisnotachieveduntilthesubstrateandco-factorsarepresentinlargeexcess.Whenassayedundertheseconditions,lightemittedfromthereactionisdirectlyproportionaltothenumberofluciferaseenzymemolecules.Thisfireflyluciferaseassaykitisdesignedforsimpleandefficientquantitationoffireflyluciferasereporterenzymeactivityfromculturedcellswithhighsensitivityandlinearity. ThiskitisanewandimprovedreplacementfortheFirefly LuciferaseAssayKit(catalogno.30003).ItfeaturesFireflyAssayBuffer2.0,whichhasbetterworkingsolutionstABIlitycomparedtotheoriginalassaybuffer.
Thisisaflash-typeluminescenceassaythatrequiressignaltobemeasuredimmediatelyafteraddingworkingsolutiontosamples.BiotiumalsoofferstheSteady-LucHTSFireflyAssayKit (cat.no.30028L),whichisahomogenousglow-typeassaywithsignalhalf-lifeof3-5hours.
BiotiumalsoofferstheFirefly&RenillaLuciferaseSingleTubeAssayKit(catalogno.30081),acombinedluciferaseassaykitforsequentialmeasurementofFireflyandRenillaluciferaseactivitysequentiallyinthesamesamplewithhighsensitivityandlinearity.
Assaysizeisbasedon96-wellformat.Thekitsinclude enoughcelllysisbuffertoperformthestatednumberofassaysinplatesizesrangingfrom96-wellto24-well.Additional 5XFireflyLysisBuffer(catalogno.99923) canbepurchasedseparatelyifneeded.
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
1.Alam,J.andJ.L.Cook.1990.Reportergenes:Applicationtothestudyofmammaliangenetranscription.Anal.Biochem.188:245-254.
2.Bronstein,I.,etal.1994.Chemiluminescentandbioluminescentreportergeneassays.Anal.Biochem.219:169-181.
3.Gould,S.J.andS.Subramani.1988.FireflyluciferaseasatoolinmolecularandcellBIOLOGy.Anal.Biochem.175:5-13.
4.Brasier,A.R.,etal.1989.OptimizeduseoftheFireflyluciferaseassayasareportergeneinmammaliancelllines.BioTechniques.7:1116-1122.
