ThiskitisdesignedtomeasureRenillaluciferaseactivityintransfectedcellreportergeneassays.Renillaluciferasehasbeenusedasareportergeneforstudyinggeneregulationandfunctioninvitroandinvivo. Itcommonlyisusedinmultiplextranscriptionalreporterassaysorasanormalizingtransfectioncontrolforfireflyluciferaseassays.Renillaluciferase,amonomeric36,000Daltonprotein,catalyzescoelenterazineoxidationbyoxygentoproducelight.Theenzymedoesnotrequirepost-translationalmodificationforitsactivityandmayfunctionasageneticreporterimmediatelyfollowingtranslation.Coelenterazinealsoemitslightfromenzyme-independentoxidation,aprocessknownasautoluminescence.Theautoluminescenceisenhancedbysuperoxideanionandperoxynitriteincellsandtissues.Thisassaykitutilizesaspecialbufferformulationdesignedtoyieldreliable,linearmeasurementsofRenillaluciferaseactivitywithminimalautoluminescencebackgroundandsuperiorsensitivity.Thiskitisaflash-typeluminescenceassay,whichrequiresluminescencetobemeasuredimmediatelyafteraddingreagentstoyoursample.
ThiskitisanewandimprovedreplacementfortheRenillaLuciferaseAssayKit(catalogno.30004).Inadditiontoprovidingasimplifiedprotocol,theRenillaLuciferaseAssayKit2.0featuresBiotium’sAquaphile™Coelenterazine,awatersolublesubstratethatcanbestoredat-20°Cwithminimalevaporation,unlikemethanolsolutionsofcoelenterazine.
Thestatednumberofassaysisbasedon96-wellassayformat.ThekitsincludeenoughPassiveLysisBuffertoperformthestatednumberofassaysinplatesizesrangingfrom96-wellto24-well.Ifneeded,additional5XPassiveLysisBuffer(catalogno.99912)canbepurchasedseparately(seebelow).
BiotiumalsoofferstheFirefly&RenillaLuciferaseSingleTubeAssayKit(catalogno.30081),acombinedluciferaseassaykitforsequentialmeasurementofFireflyandRenillaluciferaseactivitysequentiallyinthesamesamplewithhighsensitivityandlinearity.
Kitcomponents
- 5XPassiveLysisBuffer
- RenillaLuciferaseAssayBuffer2.0
- Aquaphile™Coelenterazine
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.BhaumikS.etal.(2004)JBiomedOpt.9,578-86.
2.MatijasevicZ.etal.(2001)Carcinogenesis.22,661-4.
3.NieuwenhuijsenBW.etal.(2004)JBiomolScreen.8,676-84.
4.Matthews,J.C.,Hori,K.andCormier,M.J.(1977)Biochemistry16,85–91.
