ProductHighlight

• Basedontheuniquemousemonoclonalanti-cGMPantibody
• Theonlyassaykitonthemarketwithmonoclonalanti-cGMPantibody(allotherkitsarebasedonpolyclonalanti-cGMPantibodies)
• Themonoclonalanti-cGMPantibodydisplaysmuchhigherselectivityovercAMPandGTP,thanpolyclonalanti-cGMPantibody
• Avoidsthebatch-to-batchvariationsassociatedwithpolyclonalantibodyproductionsfromanimals,thusprovidingthereproducibility

ProductDescription

Guanosine3",5"-cyclicmonophosphate(cyclicGMP;cGMP)playscriticalregulatoryrolesinmanyphysiologicalprocesses.cGMPisproducedfromGTPbyguanylylcyclasesandisdegradedbyphosphodiesterases.StimulationofguanylylcyclasesorinhibitionofphosphodiesterasescanincreasecellularcGMPconcentrations.InhibitorsofthecGMP-specificphosphodiesterasesareusedfortreatinghumandiseases.Forexample,inhibitorsofcGMPspecificphosphodiesterasetype5(suchasViagraandCialis)areusedfortreatingerectiledysfunction.

Toscreenforinhibitorsofphosphodiesterasesorstimulatorsofguanylylcyclases,itisessentialtohaveasensitive,selectiveandreproducIBLemethodtomeasurethecGMPconcentrations.Thisisespeciallytruefortheinitialscreeningsgiventhepossibleweakereffectsoflargerpoolsofcompounds.

CurrentlyavailableotherELISAkitsmeasuringcGMPlevelsarebasedonthenon-affinity-purifiedpolyclonalanti-cGMPantibody.Despitetheclaimedselectivity,thesepolyclonalanti-cGMPantibodiesdisplaycertaincross-reactivitywithcAMPorGTP.Inmostcelltypes,cGMPispresentatlevels~100foldlowerthancAMP.

NewEastBiosciencescGMPELISAkitisbasedontheuniquemousemonoclonalanti-cGMPantibody.Thismonoclonalanti-cGMPantibodydisplays>10⁸foldofselectivityovercAMP,GTPandothernucleosideanalogues.NewEastBiosciencescGMPELISAkitprovidessignificantlyimprovedsensitivityandselectivityoverotherkitsbasedonpolyclonalanti-cGMPantibodies.Ourmonoclonalanti-cGMPantibody-basedELISAkitalsoavoidsthebatch-to-batchvariationsassociatedwithpolyclonalantibodyproductionsfromanimals,thusprovidingthereproducibilityinthelongrun.

PrincipleOutline

NewEastBiosciencescGMPELISAKitisacompetitiveimmunoassaytomeasurethecGMPlevels,eitherfromcellextractsorfrominvitroguanylylcyclaseassays.Briefly,multi-wellplatesarecoatedwithgoat-anti-mouseserum.cGMPincellextractsorininvitroguanylylcyclaseassayswillcompetitivelybindtothemonoclonalanti-cGMPantibodyinthepresenceoffixedamountsofcGMP-conjugatedhorse-rADIshperoxidaseoralkalinephosphatase.KnownamountsofcGMPareusedasstandardstogeneratethecalculationcurve.Afterashortincubation,theexcessreagentsarewashedawayandsubstrateisadded.Themultiwellplatesarethenreadonamicroplatereaderat450nmor405nm.TheintensityoftheyellowcolorisinverselyproportionaltotheconcentrationofcGMPinsamples.ThemeasuredopticaldensityisusedtocalculatetheconcentrationofcGMPinsamplesbasedonthecurvefromthecGMPstandards.

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Publications:

1.  Stimulationofguanylylcyclase-Dbybicarbonate    Biochemistry,2009

2.  RoleofSulfhydryl-DependentDimerizationofSolubleGuanylylCyclaseInRelaxationofPorcineCoronaryArterytoNitricOxide    OxfordJournals-CardiovascularResearch,2011

3.  Molecularanalysis,developmentalfunctionandheavymetal-inducedexpressionofABCC5inzebrafish    ComparativeBiochemistryandPhysiologyPartB:BiochemistryandMolecularBIOLOGy,2010

4.  FeedbackControlThroughcGMP-DependentProteinKinaseContributestoDifferentialRegulationandCompartmentationofcGMPinRatCardiacMyocytesCirculationResearch,2010

5.  Pressure-Overload–InducedSubcellularRelocalization/OxidationofSolubleGuanylylCyclaseintheHeartModulatesEnzymeStimulationCirculationResearch.2012

6.  InhibitionofStriatalSolubleGuanylylCyclase-cGMPSignalingReversesBasalGangliaDysfunctionandAkinesiainExperimentalParkinsonismPLoSONE,2011

7.  Sulfhydryl-dependentdimerizationofsolubleguanylylcyclasemodulatestherelaxationofporcinepulmonaryarteriestonitricoxideYe,L.Liu,J.Liu,H.Ying,L.Dou,D.Chen,Z.Xu,X.Raj,J.U.Gao,Y.2012

8.  Natriureticpeptidesblocksynaptictransmissionbyactivatingphosphodiesterase2AandreducingpresynapticPKAactivityHu,F.Ren,J.Zhang,J.E.Zhong,W.Luo,M.2012

9.  Molt-inhibitinghormonefromChinesemittencrab(Eriocheirsinensis):Cloning,tissueexpressionandeffectsofrecombinantpeptideonecdysteroidsecretionofYOsZhang,Y.Sun,Y.Liu,Y.Geng,X.Wang,X.Wang,Y.Sun,J.Yang,W.2011