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    當前位置 ： Millipore >>> Millipore/MAB3424 | Anti-BrdU Antibody, clone AH4H7-1 / 131-14871/MAB3424/50 µg
    Millipore/MAB3424 | Anti-BrdU Antibody, clone AH4H7-1 / 131-14871/MAB3424/50 µg
    • Millipore/MAB3424 | Anti-BrdU Antibody, clone AH4H7-1 / 131-14871/MAB3424/50 µg

    Millipore/MAB3424 | Anti-BrdU Antibody, clone AH4H7-1 / 131-14871/MAB3424/50 µg

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    貨號: MAB3424
    品牌: Millipore
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      • Description
        CatalogueNumberMAB3424
        ReplacesMAB1467
        BrandFamilyChemicon®
        TradeName
        • Chemicon
        DescriptionAnti-BrdUAntibody,cloneAH4H7-1/131-14871
        AlternateNames
        • BrdU
        BackgroundInformationBromodeoxyuridine(BrdU)isathymidineanalogandisspecificallyincor-poratedintoDNAduringDNAsynthesis.Anti-bromodeoxyuridinemonoclonalantibodyisusedtoidentifycellsthathaveincorporatedBrdU.ThisimmunologicaldetectionschemehasseveraladvantagesovertheuseofrADIoactivethymidineincorporationforidentifyingcellsunder-goingreplication.Labelinganddetectioncanbeperformedthesamedayinsteadofwaitingseveraldays,asrequiredforautoradiographyoftritium-labeledcells,andthenecessityofusingmultiplespecimensforobtainingtheoptimalexposuretimeiseliminated.Inaddition,anti-bromodeoxyuridinestainingwithflowcytometricanalysisallowsmultipleparameterstobeevaluatedsimultaneously.Anti-bromodeoxyuridinemonoclonalantibodyhasbeenusedforidenti-fyingproliferatingcellsinblood(Campanaetal.,1988),tissues(Schutteetal.,1987;Hayashi,etal.,1988),tumors(Hoshinoetal.,1986;Morstynetal.,1983),aswellasfordeterminingplasmacelllabelingindices(Greippetal.,1985).
        ProductInformation
        FormatPurified
        Control
        • AfterincorporationofBrdU,allDNAcontainingspecies
        PresentationLiquidin10mMPhosphatebuffer,pH7.4containing150mMNaCland0.1%sodiumazide
        StorageandShippingInformation
        StorageConditionsMaintainfor6monthsat-20°Cfromdateofshipment.Aliquottoavoidrepeatedfreezingandthawing.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.
        Applications
        ApplicationUseAnti-BrdUAntibody,cloneAH4H7-1/131-14871(MouseMonoclonalAntibody)validatedinFC,ICC,IHCtodetectBromodeoxyuridinealsoknownasBrdU.
        KeyApplications
        • FlowCytometry
        • Immunocytochemistry
        • Immunohistochemistry
        ApplicationNotesImmunohistochemistry:(6μg/ml)

        FlowCytometry:(0.2μg/100μl/10E6cells)Optimalworkingdilutionsmustbedeterminedbyenduser.

        APPLICATIONS

        Flowcytometry:ThemethodbelowisbasedonthatofM.Vanderlaanetal.(1986).Variationsofthismethodexistintheliterature,oneconsiderationbeingtheeffectvariousfixationprocedureshaveonthelight-scatteringpropertiesofdifferentcellpopulations.Procedure:

        1.Tolabelcells,pulsewith10μMbromodeoxyuridinefor30minutes.Harvestcellsfromculture.

        2.Fixcellsin70%ethanolat+2-8°Cforatleast30min.ExtracthistonesbyresUSPendingcellsin1mLchilled0.1MHCIcontaining0.5%TritonX-100;incubatethesuspensiononicefor10minutes.Diluteacidwith5mLdistilledwaterandcentrifugeat200xgfor10min.Resuspendcellsin2mLdistilledwater.

        3.DenaturecellularDNAbysubmergingthecellsuspensionintoaboilingwaterbathfor10min.Afterwards,quicklycoolbyplacingthecellsuspensioninaniceslurryforseveralminutes.WashcellsinPBSthatcontains0.5%TritonX-100.

        4.Resuspendthecells(1-2x106cells)in100μLofsolutioncontainingapproximately2μg/mLanti-bromodeoxyuridineantibodydilutedinPBScontaining0.1%BSA(0.2μg/test).Incubatefor30minatroomtemperature.WashcellswithPBS.

        5.Resuspendcellsin100μLofdilutedgoatanti-mouseIgG-FlTCWashcellswithPBS.

        APPLICATIONS(Cont.)

        Immunohistochemistry:BelowisaprocedureforstainingcellsthathavebeenlabeledwithBrdUinvivoorinvitro.TheprocedureisbasedonthemethodsofB.Schutteetal.(1987)andD.Campanaetal.(1988).

        Preparationoftissue:

        Injectanimalwith50mgBrdU/kgbodyweight.Sacrificeanimalonehourlaterandremoveorganortissueunderstudy.EmbedtissueinOCTmediumandsnap-freezebyimmersionintoliquidnitrogen.Cut4mmfrozensectionswithacryostat.Placesectionsoneitheralbumin-orgelatin-coatedslides.

        Preparationofcells:

        Pulsecellswith10mMBrdUfor60min.Cellsgrownoncoverslips,orcytocentrifugepreparationsmadefromcellsgrowninsuspension,canbeusedforanti-bromodeoxyuridinestainingaccordingtotheprocedurebelow.

        Procedure

        1.Fixtissuesectionsorcells(onslideorcoverglass)byimmersinginabsolutemethanolfor10minutesat+2-8°C.Airdryafterremovingfromfixative.Theslidescanbestoredat-20°Cinasealedbox,orrehydratedtopreparefortheassayprocedure.Torehydrate,immerseinPBSfor3min.

        2.DenatureDNAbyincubatingtheslidesin2NHCIfor60minat+37°C.

        3.Neutralizetheacidbyimmersingtheslidesin0.1Mboratebuffer,pH8.5.Changethebuffertwiceovera10minperiod.

        4.WashslideswithPBS,changingthesolutionthreetimesovera10minperiod.

        5.Placeslidesinahumidifiedchamber(e.g.,asealedplasticboxlayeredwithwetpapertowels)andcovercellswith150-300μLofsolutioncontainingapproximately6μg/mLanti-bromodeoxyuridineantibodydilutedinPBSwith0.1%BSA.Incubatefor60minatroomtemperature.

        6.WashslideswithPBS,changingthesolutionthreetimesovera10minperiod.

        7.Applyoptimaldilutionofasecondantibodyconjugate(e.g.,anti-mouseIgG-peroxidase),incubate,wash,andperformdetectionwithasubstratethatproducesaninsolubleproduct.Afterdetection,counterstainwithHarris-modifiedhematoxylinifdesired.Slidescanthenbedehydratedandmounted.
        BIOLOGicalInformation
        ImmunogenBromodeoxyuridine-bovineserumalbuminconcentrate
        CloneAH4H7-1/131-14871
        ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
        HostMouse
        SpecificityBindstobromodeoxyuridineandcrossreactswithiodouridine(10%).Anti-bromodeoxyuridinedoesnotcrossreactwithfluorodeoxyuridine,norwithanyendogenouscellularcomponentssuchasthymidineoruridine.
        IsotypeIgG1
        SpeciesReactivity
        • All
        AntibodyTypeMonoclonalAntibody
        PurificationMethodProteinAPurfied
        MolecularWeightdependentuponthemolecularweightofthebromodeoxyuridineincorporatedproteinbeingdetected
        PhysicochemicalInformation
        Dimensions
        MaterialsInformation
        MaterialsInformation
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