首页 > BPS Bioscience > Bpsbioscience/PDE1B Cell-Based Reporter Assay Kit/60601/500 reactions

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Bpsbioscience/PDE1B Cell-Based Reporter Assay Kit/60601/500 reactions

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60601

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Catalog#:60601Size:500reactionsContactusforpricingSpecies:HumanBackground:Phosphodiesterases(PDEs)regulatetheintracellularlevelsofcAMPandcGMPbyhydrolyzingcAMPandcGMPtotheirinactive5"monophosphates.Thesecyclicnucleotidesplayanimportantroleassecondmessengersindiversephysiologicalfunctions.PDE1Bplaysaroleinsignaltransductionbyregulatingtheintracellularconcentrationofcyclicnucleotides.PDE1BhydrolyzesbothcAMPandcGMPtonucleoside5-primemonophosphate.AlthoughitpreferscGMPatlowsubstratelevels,ithasaVmaxthatisapproximatelyequalforbothcGMPandcAMP(Polli,J.W.andKoncaid,R.L.,1994;Yu,J.etal.,1997).Description:ThePDE1BCell-BasedReporterAssayKitisdesignedforidentificationofPDE1Binhibitorsinculturedcells.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.Synonym(s):PDE1B,phosphodiesterase1BSuppliedAs:ThekitincludesarecombinantHEK293celllinethatstablyexpresseshumanPDE1B,atrialnatriureticpeptide(ANP),mammalianexpressionconstructsforANPreceptorA,CNGA2,andaequorin,andbufferfortheassay.Format:Instructionsforuse:Seeassaykitdatasheetfordetailedprotocol.Storage/StABIlity:Storecelllineinliquidnitrogenimmediatelyuponreceipt.Storeothercomponentsasrecommended.Componentsarestableatleast12monthsfromdateofreceipt,whenstoredasdirected.Application(s):ThePDE1BCell-BasedReporterAssayKitisdesignedforscreeninginhibitorsofPDE1BandformonitoringPDE1B/cGMP/Ca2+channelsignalingpathwayactivityinvivo.Notes:Thevectorsaresuitablefortransienttransfection.TheyareNOTsuitablefortransformationandamplificationinbacteria.LicenseDisclosure:Purchaseofthiscelllinegrantsyouwitha10-yearlicensetousethiscelllineinyourimmediatelaboratory,forresearchuseonly.Thislicensedoesnotpermityoutoshare,distribute,sell,sublicense,orotherwisemakethecelllineavailableforusetootherlaboratories,departments,researchinstitutions,hospitals,universities,orbiotechcompanies.Thelicensedoesnotpermittheuseofthiscelllineinhumansorfortherapeuticordruguse.InappropriateuseordistributionofthiscelllinewillresultinrevocationofthelicenseandresultinanimmediateceaseofsalesanddistributionofBPSproductstoyourlaboratory.PublicationsusingthiscelllineshouldreferenceBPSBioscience,Inc.,SanDiego.Warning(s):Avoidfreeze/thawcyclesScientificCategory:PDEProductType:AssayKitDatashownislot-specific.Contactusforspecificinformationonotherlots.

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