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    當前位置 ： Millipore >>> Millipore/17-460 | EGFR STAR ELISA Kit/17-460/96 assays
    Millipore/17-460 | EGFR STAR ELISA Kit/17-460/96 assays
    • Millipore/17-460 | EGFR STAR ELISA Kit/17-460/96 assays

    Millipore/17-460 | EGFR STAR ELISA Kit/17-460/96 assays

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    貨號: 17-460
    品牌: Millipore
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      • Description
        CatalogueNumber17-460
        BrandFamilyUpstate
        TradeName
        • STAR
        • Upstate
        DescriptionEGFRSTARELISAKit
        AlternateNames
        • Epidermalgrowthfactorreceptor
        • Proto-oncogenec-ErbB-1
        • Receptortyrosine-proteinkinaseerbB-1
        • EGFReceptor
        • EGFR
        • ERBB
        • ERBB1
        BackgroundInformationI.TESTPRINCIPLE
        TheUPSTATE?colorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.TheEGFRplateiscoatedwithaspecificmousemonoclonalanti-EGFRcaptureantibody.SamplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingEGFRantigentobecapturedintheplatewells.Theplateisthenwashedtoremoveanyun-boundnon-nspecificmaterial.Aspecificrabbitanti-EGFRantibodyisaddedtodetectthecapturedfulllengthEGFRontheplatewell.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader.Thisallowsforasensitiveenzymaticdetectionofthesample.
        Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.ThekitalsoincludesastandardthatisrunasbothapositivecontrolandtotogenerateastandardcurveforEGFRmeasurement.

        II.EGFRBACKGROUND
        Epidermalgrowthfactorreceptor(EGFR)isareceptortyrosinekinasethatplaysakeyroleintheregulationofessentialnormalcellularprocessesandinthepathophysiologyofhyperproliferativediseasessuchascancer.Epidermalgrowthfactorreceptor(EGFR)isareceptortyrosinekinaseknowntobeessentialformediationofbothproliferativeandsurvivalsignalstocells.ActivationoftheEGFRsignalingpathwayhasbeenlinkedwithincreasedcellproliferation,angiogenesis,metastasisanddecreasedapoptosis.EGFRisfoundinmostsolidtumorsanditsautophosphorylationactivatestheintrinsickinaseactivitytowardheterologoussubstrates,aswellascreatingdockingsitesforadapterproteinsformultipleadapterproteinstobindandnucleatesignalingcomplexesthatactivatetheRas,PI3Kinase,andPLCpathways.
        ManyoftheEGFRmutationsaredirectlyrelatedtovariousdiseasesandarethetargetofnumerousdrugs.Forinstance,theaminoacidsubstitutionL858RisoneofseveralheterozygousmutationsthathavebeenidentifiedinNon-Small-CellLungCancer(NSCLC)patientswhohaveclinicalresponsestotheEGFRinhibitorIressa?.ThereissomeevidencethatthesemutationsresultinelevatedactivityandenhancedsensitivitytoIressa?.InpatientswithtumorsbearingIressa?-sensitivemutations,resistantsubclonescontaininganadditionalEGFRmutation,T790M,emergeinthepresenceofthedrug.IthasbeenshownexperimentallythattheT790Mmutationleadstohigh-levelfunctionalresistancetoIressa.InpatientswithtumorsbearingIressa?-sensitivemutations(eg.L858R,L861Q),resistantsubclonescontainingtheT790Mmutationemergeinthepresenceofthedrug.
        MaterialsRequiredbutNotDelivered1.Multi-channelorrepeatingPipettes
        2.Plateshaker(optional)
        3.Pipettorsandtipscapableofaccuratelymeasuring1-1000μL
        4.GraduatedSEROlogicalpipettes
        5.96-wellmicrotiterPlateReaderwith450nmfilter
        6.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata
        7.Microfugetubesforstandardandsampledilutions
        8.Mechanicalvortex
        9.1litercontainer
        10.Distilledordeionizedwater
        ProductInformation
        Components
        • CapturePlatepre-coatedwithanti-EGFRantibody:(PartNo.17-460A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
        • Anti-EGFRdetectionantibody:(PartNo.17-460B)Onebottle(11mL)ofanti-EGFRdetectionantibodycontainingsodiumazide,readytouse.
        • ELISADiluent:(PartNo.17-460C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
        • 25XELISAWashBuffer:(PartNo.17-460D)Onebottle(50mL)of25XELISAWashBuffer.
        • Anti-RabbitIgGHRPconjugate:(PartNo.17-460E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal.
        • HRPDiluent:(PartNo.17-460F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal.
        • TMBSolution:(PartNo.17-460G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
        • StopSolution:(PartNo.17-460H)Onebottle(25mL)ofstopsolution,readytouse.
        • EGFRStandard:(PartNo.17-460I)TwovialsofEGFR(fulllength)standard,lyophilized.
        • PlateCovers:Twoplatecovers.
        DetectionmethodChromogenic
        StorageandShippingInformation
        StorageConditionsMaintaintheunopenedkitat2-8°Cuntilexpirationdate.
        Applications
        KeyApplications
        • ELISA
        BIOLOGicalInformation
        SpeciesReactivity
        • Human
        AnalytesAvailable
        • EGFR
        EntrezGeneNumber
        GeneSymbol
        • EGFR
        • ERBB1
        • ERBB
        • mENA
        UniProtNumber
        UniProtSummaryFUNCTION:SwissProt:P00533#Isoform2/truncatedisoformmayactasanantagoNIST.
        SIZE:1210aminoacids;134277Da
        SUBUNIT:BindsRIPK1.CBLinteractswiththeautophosphorylatedC-terminaltailoftheEGFreceptor.PartofacomplexwithERBB2andeitherPIK3C2AorPIK3C2B.TheautophosphorylatedforminteractswithPIK3C2B,maybeindirectly.InteractswithPELP1.
        SUBCELLULARLOCATION:Cellmembrane;Single-passtypeImembraneprotein.&Isoform2:Secreted.
        TISSUESPECIFICITY:Expressedinplacenta.Isoform2isalsoexpressedinovariancancers.
        PTM:PhosphorylationofSer-695ispartialandoccursonlyifThr-693isphosphorylated.&MonoubiquitinatedandpolyubiquitinateduponEGFstimulation;whichdoesnotaffecttyrosinekinaseactivityorsignalingcapacitybutmayplayaroleinlysosomaltargeting.Polyubiquitinlinkageismainlythrough"Lys-63",butlinkagethrough"Lys-48","Lys-11"and"Lys-29"alsooccur.DISEASE:SwissProt:P00533#DefectsinEGFRareassociatedwithlungcancer[MIM:211980].
        SIMILARITY:SwissProt:P00533##Belongstotheproteinkinasesuperfamily.Tyrproteinkinasefamily.EGFreceptorsubfamily.&Contains1proteinkinasedomain.
        MISCELLANEOUS:BindingofEGFtothereceptorleadstodimerization,internalizationoftheEGF-receptorcomplex,inductionofthetyrosinekinaseactivity,stimulationofcellDNAsynthesis,andcellproliferation.
        PhysicochemicalInformation
        Sensitivity
        • Sensitivity:0.1ng/mL
          RangeofDetection:0.16to10ng/mL
          SpeciesReactivity:Human
        Dimensions
        MaterialsInformation
        MaterialsInformation
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