ProductHighlight

• BasedontheuniquemousemonoclonalAnti-cAMPantibodyItistheonlyEIAkitonthemarketwithmonoclonalanti-cAMPantibody(allotherkitsarebasedonpolyclonalanti-cAMPantibodies). 
• Nomorehazardouschemical(aceticanhydride)foryourcAMPassayThemonoclonalAnti-cAMPantibodyhassimilaraffinitiesfornon-acetylatedcAMPandacetylatedcAMP.Thusacetylationwiththeaceticanhydrideispermanentlyeliminatedfromtheassay. 
• SupersensitivityandselectivityThemonoclonalAnti-cAMPantibodydisplays>10⁸foldofselectivityovercGMP,ATP,andothernucleosideanalogues.TheAnti-cAMPEIAKitprovidessignificantlyimprovedsensitivityandselectivityoverotherkitsbasedonpolyclonalanti-cAMPantibodiesonthemarket. 
• Shortestassaytimeandhigh-throughputformatTheAnti-cAMPEIAKitisthebestchoicefordrugscreenings.

ProductDescription

Adenosine3",5"-cyclicmonophosphate(cyclicAMP;cAMP)modulatesvariousphysiologicalfunctionssuchascardiovascularBIOLOGy,learningandmemory,olfaction,immuneresponse,asthmaandkidneyfunction(1,2).cAMPisproducedfromATPbyadenylylcyclasesandisdegradedbyphosphodiesterases.StimulationofadenylylcyclasesorinhibitionofphosphodiesterasescanincreasecellularcAMPconcentrations.Blockersofadenylylcyclase-activatingreceptorsandinhibitorsofthecAMP-specificphosphodiesterasesareusedfortreatinghumandiseases.Forexample,blockingagentsforcAMP-increasingbeta-adrenergicreceptors(beta-blockers)areusedfortreatingabnormalheartrhythms,highbloodpressure(hypertension),myocardialinfarctionandheartfailure.InhibitorsofcAMPspecificphosphodiesterasetypes2and4arebeingtestedforcognitionenhancement.

ToscreenforinhibitorsorstimulatorsofcellularcAMPlevels,itisessentialtohaveasensitive,selectiveandreproducIBLemethodtomeasurethecAMPconcentrations.Thisisespeciallytruefortheinitialscreeningsgiventhepossibleweakereffectsoflargerpoolsofcompounds.

CurrentlyavailableotherELISAkitsmeasuringcAMPlevelsarebasedonthenon-affinity-purifiedpolyclonalanti-cAMPantibody.Despitetheclaimedselectivity,thesepolyclonalanti-cAMPantibodiesdisplaycertaincross-reactivitywithATP.GiventhatATPisthesubstrateforthecAMPproduction,itisverydesirabletohaveanantibodywithhighspecificitytowardscAMPoverATP.

NewEastBiosciencescAMPELISAkitisbasedontheuniquemousemonoclonalanti-cAMPantibody.Thismonoclonalanti-cAMPantibodydisplays>10⁸foldofselectivityoverATP,cGMP,andothernucleosideanalogues.NewEastBiosciencescAMPELISAkitprovidessignificantlyimprovedsensitivityandselectivityoverotherkitsbasedonpolyclonalanti-cAMPantibodies.Ourmonoclonalanti-cAMPantibody-basedELISAkitalsoavoidsthebatch-to-batchvariationsassociatedwithpolyclonalantibodyproductionsfromanimals,thusprovidingthereproducibilityinthelongrun.

FurThermore,whilepolyclonalanti-cAMPantibodiesusedinotherELISAkitshavehigheraffinityforacetylatedcAMPthannon-acetylatedcAMP,NewEastBiosciencesmonoclonalanti-cAMPantibodyhassimilaraffinitiestonon-acetylatedandacetylatedcAMPmolecules.Therefore,acetylationtreatmentsofsamplesandstandardsarenotneededinNewEastBiosciencescAMPELISAkit.Thissignificantlyreducesthetimefortheassay.Theavoidanceoforganicreagentsusedintheacetylationprocessprovidesasafeandhealthyworkenvironment.

PrincipleOutline

NewEastBiosciencescAMPELISAKitisacompetitiveimmunoassaytomeasurethecAMPlevels,eitherfromcellextractsorfrominvitroadenylylcyclaseassays.Briefly,multi-wellplatesarecoatedwithgoat-anti-mouseserum.cAMPincellextractsorininvitroadenylylcyclaseassayswillcompetitivelybindtothemonoclonalanti-cAMPantibodyinthepresenceoffixedamountsofcAMP-conjugatedhorse-rADIshperoxidaseoralkalinephosphatase.KnownamountsofcAMPareusedasstandardstogeneratethecalculationcurve.Afterashortincubation,theexcessreagentsarewashedawayandsubstrateisadded.Themultiwellplatesarethenreadonamicroplatereaderat450nmor405nm.TheintensityoftheyellowcolorisinverselyproportionaltotheconcentrationofcAMPinsamples.ThemeasuredopticaldensityisusedtocalculatetheconcentrationofcAMPinsamplesbasedonthecurvefromthecAMPstandards.

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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.  Heterogeneityinrelaxationofdifferentsizedporcinecoronaryarteriestonitrovasodilators:roleofPKGandMYPT1    PflugersArch.2012Feb;463(2):257-68

2.  Antiarrhythmiceffectofprolongedmorphineexposureisaccompaniedbyalteredmyocardialadenylylcyclasesignalinginrats    PharmacolRep.2012;64(2):351-9

3.  UncouplingofM1muscarinicreceptor/G-proteininteractionbyamyloidbeta(1-42)    Neuropharmacology.2013Apr;67:272-83

4.  SubendocardialIncreaseinReactiveOxygenSpeciesProductionAffectsRegionalContractileFunctioninIschemicHeartFailure    AntioxidRedoxSignal.2013Mar20;18(9):1009-20

5.  Heterogeneityinrelaxationofdifferentsizedporcinecoronaryarteriestonitrovasodilators:roleofPKGandMYPT1    PflugersArch.2012Feb;463(2):257-68

6.  Structuralbasisofanthraxedemafactorneutralizationbyaneutralizingantibody    BiochemicalandBiophysicalResearchCommunicationsVolume417,Issue1,6January2012,Pages324–329

7.  TransgenicrescueofdefectiveCd36enhancesmyocardialadenylylcyclasesignalinginspontaneouslyhypertensiverats    PflügersArchiv-EuropeanJournalofPhysiologyOctober2013,Volume465,Issue10,pp1477-1486

8.  OpposingHDAC4nuclearfluxesduetophosphorylationbyβadrenergicactivatedPKAorbyactivityorEpacactivatedCaMKIIinskeletalmusclefibres    TheJournalofPhysiologyVolume591,Issue14,pages3605–3623,July2013

9.  SexisamajordeterminantofneuronaldysfunctioninNeurofibromatosisType1    AnnalsofNeurologyVolume75,Issue2,pages309–316,February2014

10.  cAMP–PKAinhibitionofSK3channelreducedbothCa2+entryandcancercellmigrationbyregulationofSK3–Orai1complex    PflügersArchiv-EuropeanJournalofPhysiologyOctober2014,Volume466,Issue10,pp1921-1932

11.  Immunomodulatingeffectsofcasein-derivedpeptideQEPVLandQEPVonlymphocytesinvitroandinvivo    FoodFunct.,2014,5,2061-2069

12.  AgoNIST-DependentAnd-IndependentDopamine-1-LikeReceptorSignallingDifferentlyRegulatesDownstreamEffectors    FEBSJournalVolume281,Issue21,pages4792–4804,November2014

13.  Alterationofvascularreactivityinheartfailure:Roleofphosphodiesterasestype3and4    BritishJournalofPharmacologyVolume171,Issue23,pages5361–5375,December2014

14.  Ischemia/Reperfusion-InducedCHOPExpressionPromotesApoptosisandImpairsRenalFunctionRecovery:TheRoleofAcidosisandGPR4    PLoSOne.2014Oct24;9(10):e110944

15.  Comparativestudyofsomatostatin-humanserumalbuminfusionproteinsandnaturalsomatostatinonreceptorbinding,internalizationandactivation    PLoSOne.2014Feb27;9(2):e89932