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