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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   AssayLayoutSheet 定制肽合成NewEast Biosciences为全球生物研究界提供高质量的粗制和纯化肽或拟肽。我们使用先进的自动，半自动和手动合成器以及Fmoc技术合成肽。肽酰胺键通过HOAT / HATU偶联形成。我们的肽通过制备型HPLC纯化，并且每个级分均通过使用不同溶剂系统的分析型HPLC进行进一步分析。收集并冻干仅通过质谱法验证的具有单个峰的级分。肽通常在1-2周内递送。粗肽也可以大量节省（参见价格表）。周转时间为5-7天。除了正常的肽合成外，我们还合成修饰的肽和多种抗原性肽（MAPs）。修饰在C末端，N末端和内部残基处进行。在C末端，氨基可与羧酸，异氰酸酯和磺酰氯反应形成酰胺，脲和磺酰胺。在此终端进行经典的生物素化，荧光标记和脂质化。这些修饰以及磷酸化和糖基化也可以在内部残基处进行。相反，在C末端的修饰非常具有挑战性，需要溶液阶段的干预。为了在C端进行修饰，必须将肽从树脂中释放出来。然后，用相应的化学方法修饰释放的肽。但是，我们已经开发了适当的化学方法，可以在固相的C末端引入生物素，荧光染料和胺。随着化学，我们降低了合成肽以及昂贵试剂的成本。作为NewEast的客户，我们将节省下来的钱转嫁给您。

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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