NewEast/Monoclonal Anti-cGMP EIA Kit without Acetylation [80104][5x96 well]/80104

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¥31900.00
货号:80104
浏览量:127
品牌:NewEastBio
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商品描述

ProductHighlight

  • BasedontheuniquemousemonoclonalAnti-cGMPantibodyItistheonlyEIAkitonthemarketwithmonoclonalanti-cGMPantibody(allotherkitsarebasedonpolyclonalanti-cGMPantibodies). 
  • Nomorehazardouschemical(aceticanhydride)foryourcGMPassayThemonoclonalAnti-cGMPantibodyhassimilaraffinitiesfornon-acetylatedcGMPandacetylatedcGMP.Thusacetylationwiththeaceticanhydrideispermanentlyeliminatedfromtheassay. 
  • SupersensitivityandselectivityThemonoclonalAnti-cGMPantibodydisplays>108foldofselectivityovercAMP,GTP,andothernucleosideanalogues.TheAnti-cGMPEIAKitprovidessignificantlyimprovedsensitivityandselectivityoverotherkitsbasedonpolyclonalanti-cGMPantibodiesonthemarket. 
  • Shortestassaytimeandhigh-throughputformatTheAnti-cGMPEIAKitisthebestchoicefordrugscreenings.

ProductDescription

    Guanosine3",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.

    PrincipleOutline

      NewEastBiosciencescGMPELISAKitisacompetitiveimmunoassaytomeasurethecGMPlevels,eitherfromcellextractsorfrominvitroguanylylcyclaseassays.Briefly,multi-wellplatesarecoatedwithgoat-anti-mouseserum.cGMPincellextractsorininvitroguanylylcyclaseassayswillcompetitivelybindtothemonoclonalanti-cGMPantibodyinthepresenceoffixedamountsofcGMP-conjugatedhorse-rADIshperoxidase.KnownamountsofcGMPareusedasstandardstogeneratethecalculationcurve.Afterashortincubation,theexcessreagentsarewashedawayandsubstrateisadded.Themultiwellplatesarethenreadonamicroplatereaderat450nm.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.  Sulfhydryl-dependentdimerizationofsolubleguanylylcyclasemodulatestherelaxationofporcinepulmonaryarteriestonitricoxide    PflugersArch.2013Feb;465(2):333-41
    2.  Pressure-overload-inducedsubcellularrelocalization/oxidationofsolubleguanylylcyclaseintheheartmodulatesenzymestimulation    CircRes.2012Jan20;110(2):295-303
    3.  Natriureticpeptidesblocksynaptictransmissionbyactivatingphosphodiesterase2AandreducingpresynapticPKAactivity    ProcNatlAcadSciUSA.2012Oct23;109(43):17681-6
    4.  Roleofsulfhydryl-dependentdimerizationofsolubleguanylylcyclaseinrelaxationofporcinecoronaryarterytonitricoxide    CardiovascRes.2011Jun1;90(3):565-72
    5.  Molt-inhibitinghormonefromChinesemittencrab(Eriocheirsinensis):Cloning,tissueexpressionandeffectsofrecombinantpeptideonecdysteroidsecretionofYOs    GenCompEndocrinol.2011Sep15;173(3):467-74
    6.  InhibitionofStriatalSolubleGuanylylCyclase-cGMPSignalingReversesBasalGangliaDysfunctionandAkinesiainExperimentalParkinsonism    PLoSOne.2011;6(11):e27187
    7.  Molecularanalysis,developmentalfunctionandheavymetal-inducedexpressionofABCC5inzebrafish    CompBiochemPhysiolBBiochemMolBiol.2011Jan;158(1):46-55
    8.  FeedbackcontrolthroughcGMP-dependentproteinkinasecontributestodifferentialregulationandcompartmentationofcGMPinratcardiacmyocytes    CircRes.2010Nov12;107(10):1232-40
    9.  TheSolubleGuanylateCyclaseActivatorBAY58-2667ProtectsagainstMorbidityandMortalityinEndotoxicShockbyRecouplingOrganSystems    PLoSOne.2013Aug28;8(8):e72155
    10.  Stimulationofguanylylcyclase-Dbybicarbonate    Biochemistry.2009May26;48(20):4417-22
    11.  Volumeoverloadinducesdifferentialspatiotemporalregulationofmyocardialsolubleguanylylcyclaseineccentrichypertrophyandheartfailure    JournalofMolecularandCellularCardiologyVolume60,July2013,Pages72–83
    12.  DefiningSpecificityDeterminantsofCyclicGMP-MediatedGustatorySensoryTransductioninCaenorhaBDitiselegans    genetics.113.152660v1194/4/885
    13.  Theroleofnitricoxidesignalinginfoodintake;insightsfromtheinnermitochondrialmembranepeptidase2mutantmice    RedoxBIOLOGyVolume1,Issue1,2013,Pages498–507
    14.  CrossregulationbetweencGMP-dependentproteinkinaseandAktinvasodilatationofporcinepulmonaryartery    JournalofCardiovascularPharmacology:November2014-Volume64-Issue5-p452-459
    15.  Geneticablationofsolutecarrierfamily7a3aleadstohepaticsteatosisinzebrafishduringfasting    HepatologyVolume60,Issue6,pages1929–1941,December2014