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    當前位置 ： Millipore >>> Millipore/NS170 | Phosphorylated Neurofilament (pNF-H) Sandwich ELISA Kit/NS170/1 kit96 wells
    Millipore/NS170 | Phosphorylated Neurofilament (pNF-H) Sandwich ELISA Kit/NS170/1 kit96 wells
    • Millipore/NS170 | Phosphorylated Neurofilament (pNF-H) Sandwich ELISA Kit/NS170/1 kit96 wells

    Millipore/NS170 | Phosphorylated Neurofilament (pNF-H) Sandwich ELISA Kit/NS170/1 kit96 wells

    價格: ￥6972.00 市場價: 11620.00
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    貨號: NS170
    品牌: Millipore
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      • Description
        CatalogueNumberNS170
        BrandFamilyChemicon®
        TradeName
        • Chemicon
        DescriptionPhosphorylatedNeurofilament(pNF-H)SandwichELISAKit
        BackgroundInformationTheChemiconPhosphorylatedNeurofilamentH(pNF-H)kitisasandwichenzymeimmunoassay(EIA),whichmeasurespNF-H.ThekitwillmeasurepNF-Hfromavarietyofmammalianspeciesincludinghuman,rat,mouse,bovine,porcine,andfeline.

        Neurofilamentsarethe10nmdiameterfilamentswhichmakeupthemajorpartoftheCytoskeletonofneuronsandareparticularlyconcentratedinlargerdiameteraxons.Theybelongtotheintermediateor10nmfilamentprotein/genesuperfamilywhichalsoincludesthekeratins,themajorstructuralproteinsofskin.NeurofilamentsaregenerallyregardedasbeingcomposedofthreemajorproteinsubunitsreferredtoasNF-L,NF-MandNF-H.NF-Listhe"low"or"light"subunit,NF-Misthe"medium"or"middle"subunitandNF-Histhe"heavy"or"high"subunit.ThisnomenclatureisbasedontheapparentsizeoftheseproteinsonSDS-PAGEgels;NF-Listypicallyabout68kDa,NF-Misabout150kDaandNF-Hisabout200kDa.Thesethreeproteins,referredtoastheneurofilamenttriplet,werediscoveredinthe1970sasaresultofstudiesofaxonallytransportedproteins(HoffmanandLasek1975).Afourthprotein,α-internexin,a.k.aNF66,waslaterdiscoveredasaproteinco-purifiingwiththeneurofilamenttripletproteins(PachterandLiem1985).Afifthprotein,peripherin,isfoundinsomeneurofilamentsalongwiththeotherfourproteins,mostlyintheperipheralnervoussystem(Portieretal.1983).Neurofilamentsaremajorproteinsofneuronsandareparticularlyconcentratedinaxons.TheproteinNF-Hhassomeveryunusualproperties.Firstlyitcontains50-60backtobackhexa,heptaoroctapeptiderepeatseachcontainingthesequenceLysine-Serine-Proline(KSP).Theserineresiduesinthesepeptiderepeatsarephosporylatedinvivo,andinaxonalneurofilaments,thesesitesareheavilyphosphorylated.ThephosphorylatedformsofNF-Harealsoquiteresistanttoproteases,suggestingthatthisveryabundantproteinmightbeparticularlyresistanttoproteotyticcleavageuponreleasefromdamagedanddiseasedaxons.Thismeansthatdetectionofthisproteinincerebrospinalfluid(CSF)andbloodwouldprovideinformationaboutthedegreeofaxonalinjurywhichhasoccurred(reviewedinPetzold2005).Asrecentlyshown,thisproteincanbedetectedinquitelargeamountsfollowingexperimentalspinalcordandbraininjuryinrats(Shawetal.2005).Levelsofgreaterthan100ng/mlofpNF-Hweredetectableinbloodfollowingseriousspinalcordinjuryandlowerbutstilleasilydetectablelevelswereseeninbloodofanimalsgivenexperimentalbraininjury.Morerecentstudieshaverevealedconsiderableamountsofthisproteininthebloodofmicetransgenicformutationsofhumancopper/zincsuperoxidedismutase1whichareassociatedwithamyotrophiclateralsclerosis(ALS).ThesemicedevelopanaxonaldegenerationpathologysimilartothatseeninhumanswithALS,andbloodpNF-Hlevelscanbeusedtomonitorthis.Interestingly,pNF-Hisdetectablebeforetheonsetofobviousdiseasesymptoms.OtherstudiesshowthatpNF-Hcanbedetectedintissuehomogenates.OtherexperimentsshowthatpNF-Hcanbedetectedintheplasmaofhumanssufferingfromopticneuritis(Petzoldetal2004)andinelevatedlevelsinthecerebrospinalfluid(CSF)ofindividualssufferingfrombraintumorsandstroke(PetzoldandShaw,unpublished).

        TestPrincipleWiththeChemiconpNF-Hassaysystem,chickenpolyclonalantibodiesgeneratedagainstpNF-H,arepre-coatedontoa96-wellplateandareusedtocapturepNF-Hfromasample.CapturedpNF-HisdetectedusingpNF-Hspecificrabbitpolyclonalantibodiesandagoatanti-rabbitalkalinephosphataseconjugate.AfteradditionofthesubstratesolutiontheamountofpNF-Hisdetermined.ThestandardcurvedemonstratesadirectrelationshipbetweenOpticalDensity(OD)andpNF-Hconcentration:i.e.,thehighertheODthehigherthepNF-Hconcentrationinthesample.

        ApplicationTheChemiconpNF-HkitisdesignedtomeasuretheamountofpNF-HintissuehomogenatesandBIOLOGicalfluid(serum,plasma,andserum-free)samplesfrommouseandrat.Thereareenoughreagentsincludedinthiskitforone96-wellimmuno-assayplate.Runningduplicatewellsforsamplesandstandardsisrecommended.
        MaterialsRequiredbutNotDelivered1.Multi-channelorrepeatingPipettes
        2.Pipettors&tipscapableofaccuratelymeasuring10-1000μL
        3.GraduatedSEROlogicalpipettes
        4.96-wellmicroplatereadercapableofmeasuringabsorbanceat405nm
        5.Graphpaperformanualplottingofdata
        6.1.5mLtubes
        7.Mechanicalvortex
        8.Two1litercontainers
        9.Plateshaker(optional)
        10.3NNaOH(optional)
        11.Distilledordeionizedwater.
        ProductInformation
        Components
        • ImmunoplatePre-coatedwithChickenanti-pNF-HPolyclonalAntibody:(PartNo.2006263).One96-wellimmunoplatepre-coatedwithchickenanti-pNF-Hpolyclonalantibodysealedinafoilpouch.
        • Rabbitanti-pNF-HPolyclonalAntibody:(PartNo.2006919).Onevial(100μL)ofrabbitanti-pNF-Hpolyclonalantibody.
        • Goatanti-Rabbit,AlkalinePhosphataseConjugatedAntibody:(PartNo.2006917).Onevial(10μL)ofgoatanti-rabbitalkalinephosphataseconjugate.
        • BovinepNF-HStandard:(PartNo.2006918).Onevial(5.0ng)ofbovinepNF-Hstandard.
        • 20XTBS:(PartNo.2004913).Onebottle(50mL)of20XTBS.
        • 20%Tween-20?:(PartNo.2004915).Onevial(3mL)of20%Tween-20?.
        • ElisaAssayDiluent:(PartNo.2006921)Onebottle(30mL)ofElisaassaydiluentcontaing0.05%Kathonasapreservative,providedinareadytouseformat.
        • pNPPBuffer:(PartNo.2006261).Onebottle(12mL)ofpNPPbufferforusewith50Xp-NPP,providedinareadytouseformat.
        • 50XpNPP:(PartNo.2006262).Onevial(250μL)of50XpNPP(p-nitrophenylphosphatase)alkalinephosphatasesubstrate.
        • PlateCovers-TwoPlateCovers.
        DetectionmethodChromogenic
        StorageandShippingInformation
        StorageConditionsPriortousemaintainthekitcomponentsat2°-8°Cand-20°C,asindicated,untilexpirationdate.

        1.WashBuffer-AfterdilutionmaintaintheWashSolutionat2°-8°Cforuptoonemonthoruntilexpirationdate.

        2.Tween-20?-Storeatroomtemperatureuponreceiptofkit.

        3.ElisaAssayBuffer-Aliquotasrequiredtoavoidrepeatedfreeze-thawcycles.Afterthawing,maintaintheElisaAssayBufferat2°-8°Cforuptoonemonth,oruntilexpirationdate.

        4.ImmunoplatePre-coatedwithChickenanti-pNF-HPolyclonalAntibody-Unusedstripwellsmaybereturnedtothefoilpouchandstoredat2°-8°Cforuptoonemonth,oruntilexpirationdate.
        Applications
        ApplicationThePhosphorylatedNeurofilamentH(pNF-H)kitisasandwichenzymeimmunoassay(EIA),whichmeasurespNF-H.
        KeyApplications
        • ELISA
        BiologicalInformation
        SpeciesReactivity
        • Bovine
        • Human
        • Mouse
        • Pig
        • Rat
        • Feline
        AnalytesAvailable
        • PhosphorylatedNeurofilament(pNF-H)
        EntrezGeneNumber
        GeneSymbol
        • NEFH
        • NF-H
        • NFH
        • KIAA0845
        Modifications
        • Phosphorylation
        UniProtNumber
        UniProtSummaryFUNCTION:SwissProt:P12036#Neurofilamentsusuallycontainthreeintermediatefilamentproteins:L,M,andHwhichareinvolvedinthemaintenanceofneuronalcaliber.NF-HhasanimportantfunctioninmatureaxonsthatisnotsubservedbythetwosmallerNFproteins.
        SIZE:1026aminoacids;112480Da
        PTM:ThereareanumberofrepeatsofthetripeptideK-S-P,NFHisphosphorylatedonanumberoftheserinesinthismotif.ItisthoughtthatphosphorylationofNFHresultsintheformationofinterfilamentcrossbridgesthatareimportantinthemaintenanceofaxonalcaliber.&Phosphorylationseemstoplayamajorroleinthefunctioningofthelargerneurofilamentpolypeptides(NF-MandNF-H),thelevelsofphosphorylationbeingaltereddevelopmentallyandcoincidentwithachangeintheneurofilamentfunction.
        DISEASE:SwissProt:P12036#DefectsinNEFHareacauseofsusceptibilitytoamyotrophiclateralsclerosis(ALS)[MIM:105400].ALSisaneurodegenerativedisorderaffectingupperandlowermotorneurons,andresultinginfatalparalysis.Sensoryabnormalitiesareabsent.Deathusuallyoccurswithin2to5years.Theetiologyislikelytobemultifactorial,involvingbothgeneticandenvironmentalfactors.
        SIMILARITY:SwissProt:P12036##Belongstotheintermediatefilamentfamily.
        PhysicochemicalInformation
        Sensitivity
        • Sensitivity:0.0585ng/mL
          RangeofDetection:0.0293ng/mLto15ng/mL
        Dimensions
        MaterialsInformation
        MaterialsInformation
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