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    當前位置 ： Millipore >>> Millipore/MAB5256 | Anti-Neurofilament 200 kDa Antibody, clone NE14/MAB5256/40 µg
    Millipore/MAB5256 | Anti-Neurofilament 200 kDa Antibody, clone NE14/MAB5256/40 µg
    • Millipore/MAB5256 | Anti-Neurofilament 200 kDa Antibody, clone NE14/MAB5256/40 µg

    Millipore/MAB5256 | Anti-Neurofilament 200 kDa Antibody, clone NE14/MAB5256/40 µg

    價格: ￥4440.00 市場價: 7400.00
    打印產品信息
    貨號: MAB5256
    品牌: Millipore
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      • Description
        CatalogueNumberMAB5256
        BrandFamilyChemicon®
        TradeName
        • Chemicon
        DescriptionAnti-Neurofilament200kDaAntibody,cloneNE14
        BackgroundInformationNeurofilamentsareatypeofintermediatefilamentthatserveasmajorelementsoftheCytoskeletonsupportingtheaxoncytoplasm.Theyarethemostabundantfibrillarcomponentsoftheaxon,beingonaverage3-10timesmorefrequentthanaxonalmicrotubules.Neurofilaments(10nmindia.)arebuiltfromthreeintertwinedprotofibrilswhicharethemselvescomposedoftwotetramericprotofilamentcomplexsofmonomericproteins.Theneurofilamenttripletproteins(68/70,160,and200kDa)occurinboththecentralandperipheralnervoussystemandareusuallyneuronspecific.The68/70kDaNF-Lproteincanself-assembleintoafilamentousstructure,howeverthe160kDaNF-Mand200kDaNF-Hproteinsrequirethepresenceofthe68/70kDaNF-Lproteintoco-assemble.Neuromas,ganglioneuromas,gangliogliomas,ganglioneuroblastomasandneuroblastomasstainpositivelyforneurofilaments.Althoughtypicallyrestrictedtoneurons,neurofilamentshavebeendetectedinparagangliomasandadrenalandextra-adrenalpheochromocytomas.Carcinoids,neuroendocrinecarcinomasoftheskin,andoatcellcarcinomasofthelungalsoexpressneurofilaments.FormoreneurofilamentinformationseeNervousSystemCellTypeSpecificMarkerchartonlineundertheCHEMICONTechnicalSupportsection.
        ProductInformation
        FormatPurified
        PresentationPurifiedimmunoglobulin.Liquid.Buffer=0.02MPhosphatebuffer,0.25MNaClcontaining0.1%sodiumazide.
        StorageandShippingInformation
        StorageConditionsMaintainrefrigeratedat2-8°Cinundilutedaliquotsforupto6months.DONOTFREEZE
        Applications
        ApplicationDetectNeurofilament200kDausingthisAnti-Neurofilament200kDaAntibody,cloneNE14validatedforuseinIH.
        KeyApplications
        • Immunohistochemistry
        ApplicationNotesImmunohistochemistry:5-10μg/mL(Seebelowprotocol.)

        Optimalworkingdilutionsmustbedeterminedbyenduser.

        ImmunohistochemistryProtocolforAnti-Neurofilament200kD

        Idealspecimensareobtainedfromfrozensectionsfromshock-frozentissuesamples.Thefrozensectionsaredriedintheairandthenfixedwithacetoneat-15to-25°Cfor10min.Excessacetoneisallowedtoevaporateat15-25°C.Materialfixedinalcoholandembeddedinparaffincanalsobeused,see(Altmannsbergeretal.,1982).Theantibodyappearstoreactwithtissuefixedinformaldehydeforashorttime(10min)(Debusetal.,1983).Otherfixationconditionsmustbefirsttestedbytheinvestigator.

        Itisadvantageoustoblockunspecificbindingsitesbyoverlayingthesectionswithfetalcalfserumfor20-30minat15-25°C.Excessoffetalcalfserumisremovedbydecantingbeforeapplicationoftheantibodysolution.Cytocentrifugepreparationsofsinglecellsorcellsmearsarealsofixedinacetone.Thesepreparationsshould,however,notbedriedintheair.Instead,theexcessacetoneisremovedbybrieflywashinginphosphate-bufferedsaline(PBS).

        Furthertreatmentisthenasfollows:

        Overlaythepreparationwith10-20μLantibodysolutionandincubateinahumidchamberat37°Cfor1h.

        DiptheslidebrieflyinPBSandthenwash3timesinPBSfor3min(usefreshPBSeachtime)

        Wipethemarginsofthepreparationdryandoverlaythepreparationwith10-20μLofananti-mouseIg-FITCoranti-mouseIg-PODantibodyandallowtoincubatefor1hat37°Cinahumidchamber.

        Washtheslideasdescribedabove.

        Thepreparationmustnotbeallowedtodryoutduringanyofthesteps.

        Ifusinganindirectimmunofluorescencetechnique,thepreparationshouldbeoverlaidwithasuitableembeddingmedium(e.g.Moviol,Hoechst)andexaminedunderthefluorescencemicroscope.IfaPOD-conjugatehasbeenusedasthesecondaryantibody,thepreparationshouldbeoverlaidwithasubstratesolution(seebelow)andincubatedat15-25°CuntilaclearlyvisIBLered-browncolordevelops.Anegativecontrol(e.g.onlythesecondaryantibody)shouldremainunchangedincolorduringthisincubationperiod.Subsequently,thesubstrateiswashedoffwithPBSandthepreparationisstained,ifdesired,withhemalumstainforabout1min.ThehemalumsolutioniswashedoffwithPBS,thepreparationisembeddedandexamined.

        Substratesolutions:

        Aminoethyl-carbazole:

        Dissolve2mg3-amino-9-ethylcarbazolewith1.2mLdimethylsulfoxideandadd28.8mL0.05MTris-HCl,pH7.3,and20μL3%H2O2(w/v).Preparesolutionfreshlyeachday.

        Diaminobenzidine:

        Dissolve25mg3,3"-diaminobenzidinewith50mL0.05MTris-HCl,pH7.3,andadd40μL3%H2O2(w/v).Preparesolutionfreshlyeachday.
        BIOLOGicalInformation
        ImmunogenPurifiedneurofilamentpolypeptides(Debusetal.,1983).
        CloneNE14
        ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
        HostMouse
        SpecificityTheantibodyreactswithneurofilament200kD.
        IsotypeIgG1
        SpeciesReactivity
        • Human
        • Pig
        • Rat
        AntibodyTypeMonoclonalAntibody
        EntrezGeneNumber
        GeneSymbol
        • NEFH
        • NFH
        • NF-H
        • KIAA0845
        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
        Dimensions
        MaterialsInformation
        MaterialsInformation
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