Description:

Description:PDE1Bplaysakeyroleinsignaltransductionbyregulatingtheintracellularconcentrationofcyclicnucleotides.PDE1BhydrolyzesbothcAMPandcGMPtonucleoside5?-monophosphateandpreferscGMPatlowsubstratelevels.ThePDE1BCell-BasedReporterAssayKitisdesignedforidentificationofPDE1Binhibitorsinculturedcells.ThekitincludesarecombinantHEK293celllinethatstablyexpresseshumanPDE1B,atrialnatriureticpeptide(ANP),mammalianexpressionconstructsforANPreceptorA,CNGA2,andaequorin,andbufferfortheassay.InthePDE1Breporterassay,PDE1B-HEK293cellsaretransientlytransfectedwithexpressionvectorsforatrialnatriureticpeptide(ANP)receptorA(ANPRA,aguanylatecyclase),cyclicnucleotide-gatedcationchannelCNGA2,andthephotoproteinaequorin.WhencellsareactivatedwithANP,ANPinducestheproductionofcGMPthroughANPRA.cGMPcanactivatetheCNGA2channelandinducesCa2+influx,resultinginaequorinluminescence.Therefore,theintracellularcGMPlevelismonitoredviaaequorinluminescenceinducedbyCa2+influxthroughCNGA2,actingastheintracellularcGMPsensor.PDE1BcanreducetheANP-inducedcGMPlevelinthecells,causingloweredaequorinluminescence.WhenPDE1Bactivityisinhibited(forexample,bythePDE1inhibitorBAY607550),thentheANP-inducedcGMPlevelincreases,resultingincalciuminfluxthatstimulatesluminescencesignals.Aschematicpresentationofthisassayisshowninfigure1.

Format:assaykit

ScientificCategory:PDE

Species:human

AdditionalInformation:

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Name	PDE1BCell-BasedReporterAssayKit
RelatedProductNames	PDE,phosphodiesterase,screening,inhibitors,PDE1B
Format	AssayKit
Reactivity	Human
Storage	Mixedstorage
IntendedUse	ResearchUseOnly