Description:

Description:Phosphodiesterases(PDEs)playanimportantroleinthedynamicregulationofcAMPandcGMPsignaling.PDE4Dhas3",5"-cyclic-AMPphosphodiesteraseactivityanddegradescAMP.InhibitionofPDE4DactivitybyitsinhibitorsleadstoanelevatedintracellularlevelofcAMP.ThePDE4Dgeneencodesatleast9differentisoforms,andhasbeenlinkedtostroke,asthma,arrhythmia,andcardiacmyopathy,makingitanimportanttherapeutictarget.ThePDE4Dcell-basedactivityassayisdesignedforscreeninginhibitorsofPDE4D7inculturedcells.TheassayisbasedontransfectingcellswiththeCREluciferasereporter.CREreportercontainsthefireflyluciferasegeneunderthecontrolofcAMPresponseelement(CRE).ElevationofintracellularcAMPactivatesCREbindingprotein(CREB)tobindCREandinducetheexpressionofluciferase.ForskoliniscommonlyusedtoraisetheintracellularlevelofcAMPincellphysiologystudies.WhencellstransientlytransfectedwithCREreporterareactivatedbyforskolin,theintracellularlevelofcAMPisupregulated,whichinducestheexpressionofCREluciferasereporter.However,whencellsareco-transfectedwithPDE4D7expressionvectorandCREreporter,thelevelofforskolin-inducedcAMPisreduced,resultinginlowerexpressionlevelofluciferase.WhencellsaretreatedwithPDE4DinhibitortoinhibitPDE4D7activity,cAMPlevelisrestored,resultinginhigherluciferaseactivity.ThekitincludesCREluciferasereporter(premixedwithconstitutively-expressingRenilla(seapansy)luciferasevectorthatservesasaninternalcontrolfortransfectionefficiency),PDE4D7expressionvector,andforskolin.

Format:assaykit

ScientificCategory:PDE

AdditionalInformation:

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Name	PDE4DCell-BasedActivityAssayKit
RelatedProductNames	PDE4D
Format	AssayKit
Storage	-20°C
IntendedUse	ResearchUseOnly