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ProductDescriptionGuanosine3",5"-cyclicmonophosphate(cyclicGMP;cGMP)playscriticalregulatoryrolesinmanyphysiologicalprocesses.cGMPisproducedfromGTPbyguanylylcyclasesandisdegradedbyphosphodiesterases.StimulationofguanylylcyclasesorinhibitionofphosphodiesterasescanincreasecellularcGMPconcentrations.InhibitorsofthecGMP-specificphosphodiesterasesareusedfortreatinghumandiseases.Forexample,inhibitorsofcGMPspecificphosphodiesterasetype5(suchasViagraandCialis)areusedfortreatingerectiledysfunction. Toscreenforinhibitorsofphosphodiesterasesorstimulatorsofguanylylcyclases,itisessentialtohaveafast,sensitive,selectiveandreproducIBLemethodtomeasurethecGMPconcentrations.Thisisespeciallytruefortheinitialscreeningsgiventhepossibleweakereffectsoflargerpoolsofcompounds. CurrentlyavailableotherELISAkitsmeasuringcGMPlevelsarebasedonthenon-affinity-purifiedpolyclonalanti-cGMPantibody.Despitetheclaimedselectivity,thesepolyclonalanti-cGMPantibodiesdisplaycertaincross-reactivitywithcAMPorGTP.Inmostcelltypes,cGMPispresentatlevels~100foldlowerthancAMP. NewEastBiosciencescGMPELISAkitisbasedontheuniquemousemonoclonalanti-cGMPantibody.Thismonoclonalanti-cGMPantibodydisplays>108foldofselectivityovercAMP,GTP,andothernucleosideanalogues.NewEastBiosciencescGMPELISAkitprovidessignificantlyimprovedsensitivityandselectivityoverotherkitsbasedonpolyclonalanti-cGMPantibodies.Ourmonoclonalanti-cGMPantibody-basedELISAkitalsoavoidsthebatch-to-batchvariationsassociatedwithpolyclonalantibodyproductionsfromanimals,thusprovidingthereproducibilityinthelongrun. FurThermore,whilepolyclonalanti-cGMPantibodiesusedinotherELISAkitshavehigheraffinityforacetylatedcGMPthannon-acetylatedcGMP,NewEastBiosciencesmonoclonalanti-cGMPantibodyhassimilaraffinitiestonon-acetylatedandacetylatedcGMPmolecules.Therefore,acetylationtreatmentsofsamplesandstandardsarenotneededinNewEastBiosciencescGMPELISAkit.Thissignificantlyreducesthetimefortheassay.Theavoidanceoforganicreagentsusedintheacetylationprocessprovidesasafeandhealthyworkenvironment. PrincipleOutlineNewEastBiosciencescGMPELISAKitisacompetitiveimmunoassaytomeasurethecGMPlevels,eitherfromcellextractsorfrominvitroguanylylcyclaseassays.Briefly,multi-wellplatesarecoatedwithgoat-anti-mouseserum.cGMPincellextractsorininvitroguanylylcyclaseassayswillcompetitivelybindtothemonoclonalanti-cGMPantibodyinthepresenceoffixedamountsofcGMP-conjugatedhorse-rADIshperoxidase.KnownamountsofcGMPareusedasstandardstogeneratethecalculationcurve.Afterashortincubation,theexcessreagentsarewashedawayandsubstrateisadded.Themultiwellplatesarethenreadonamicroplatereaderat450nm.TheintensityoftheyellowcolorisinverselyproportionaltotheconcentrationofcGMPinsamples.ThemeasuredopticaldensityisusedtocalculatetheconcentrationofcGMPinsamplesbasedonthecurvefromthecGMPstandards.
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