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    當前位置 ： Millipore >>> Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL
    Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL
    • Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL

    Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL

    價格: ￥3576.00 市場價: 5960.00
    打印產品信息
    貨號: MAB4078
    品牌: Millipore
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      • Description
        CatalogueNumberMAB4078
        BrandFamilyChemicon®
        TradeName
        • Chemicon
        DescriptionAnti-PCNAAntibody,clone131-11912
        AlternateNames
        • ProliferatingCellNuclearAntigen
        • DNAPolymerasedeltaProcessivityFactor
        ProductInformation
        FormatAscites
        PresentationAscitesfluidcontainingnopreservatives
        StorageandShippingInformation
        StorageConditionsMaintainfrozenat-20°Cinundilutedaliquotsforupto12monthsfromdateofreceipt.Avoidrepeatedfreeze/thawcycles.
        Applications
        ApplicationAnti-PCNAAntibody,clone131-11912isamousemonoclonalantibodyfordetectionofPCNAalsoknownasProliferatingCellNuclearAntigen,DNAPolymerasedeltaProcessivityFactor&hasbeenvalidatedinIF,ICC,IHC(P).
        KeyApplications
        • Immunofluorescence
        • Immunocytochemistry
        • Immunohistochemistry(Paraffin)
        ApplicationNotesImmunohistochemistryonformalinfixed,deparaffinizedtissuesections
        Frozentissuesections.
        Smears.
        Cytospins.
        *DonotuseSeruminblockingsteps.

        Alsoseehttp://ajp.amjpathol.org/cgi/content/abstract/141/6/1453forimportantreactivityinformationon19A2inparaffinfixedtissues.

        Finalworkingdilutionsmustbedeterminedbyenduser.

        IMMUNOHISTOCHEMICALPROCEDUREFORMAB4078

        ThefollowinghistochemicalimmunoperoxidaseprotocolhasbeenoptimizedforusewithMAB4078.

        METHODOLOGY:ThismethodisusedwithstandardstreptavidinorABCimmunoperoxidasemethodsusingaDABsubstraterunatroomtemperaturewithparaffinorfrozensections,smearsorcytospins.

        NOTE:Optimalfixationtimeforparaffinprocessedtissuefixedin10%NeutralBufferedFormalinis12-14hours.

        REQUIREDSUPPLEMENTALMATERIAL:

        1.3%HydrogenPeroxide:commerciallyavailable.

        2.PBS

        3.EthylAlcohol:histologygradeorequivalent

        4.Methanol:histologygradeorequivalent

        5.Acetone:histologygradeorequivalent

        6.Xylene(histologygrade)orXyleneSubstitute(Hemo-D)

        7.MountingMedia:Krystalon(Harleco)orequivalent

        8.Gill#1Hematoxylin:Sigma#GHS-1-32orequivalent

        9.GoatAnti-MouseBiotin

        10.StreptavidinHRP(optionalreagent)

        11.3,3"Diaminobenzidinetetrahydrochloride(DAB):Sigma#D-5637orequivalent

        12.ABC:VectorKit#PK4000orequivalent

        REAGENTPREPARATION:

        1.Primaryantibody:PrepareastocksolutionofantibodyinPBS

        2.GoatAnti-MouseBiotin:Approximatedilutionof1:100inPBS(10mg/test)

        3.Streptavidin:5mLstocksolutionand995mLPBS(1/200dilution).Letsit15minutesbeforeuse(yieldasavolumesufficientfor~20slides)ORifusingABCkitfromVectorfollowasinstructed.

        4.DAB:Reconstitute1.5mgofpowderwith100mLofPBS.Activate15minutespriortousebyadding100mL3%H2O2(yieldsavolumesufficientfor~20slides).WARNING:DABisapossIBLecarcinogen.Avoidcontactwithskin.

        5.PBS:Usingagraduatedcylinder,reconstitutewith500mLofdistilledwater.

        6.AlcoholBaths:Alldilutionsarealcoholsmadewithdistilledwater.

        PROCEDURE:

        PARAFFINSECTIONS

        (cutat4-5microns,10%NeutralBufferedFormalinfixed):

        1.Placeslidesin62°Covenfor15minutes.

        2.PlaceslidesinHemo-D,orxylene,for15minutes.

        3.De-paraffinizeslidesthroughbathsof:A)xylene/100%ethanol1:1(orHemo-D/100%ethanol1:1),B)100%ethylalcoholx2,C)95%ethylalcohol,D)90%ethylalcohol,E)80%ethylalcohol,F)70%ethylalcoholx2(2minuteseachbath).

        4.Placeslidesinrunningcooltapwaterfor5minutes.

        5.Shakeoffexcesswater.Placeslidesonincubationracksheldatroomtemperature.CirclesectionswithPAPpen(Newcomer,Inc.)ordiamondpentoisolatesection.

        6.Floodsectionswithprimaryantibodyandincubatefor35-40minutes.

        7.WipeoffexcessPBS.FloodsectionswithGoatAnti-MouseBiotinreagentandincubatefor35-40minutes.

        8.WashwithPBS3-5min

        9.WashagainwithPBS3-5min.

        10.WipeoffexcessPBS.Floodsectionswithstreptavidin-HRPandincubatefor10minutesORfloodsectionswithABCandincubatefor35-40minutes.

        11.WashwithPBS3-5min.

        12.WipeoffexcessPBS.FloodsectionswithDABandincubatefor6-8minutes.

        13.Placeslidesincoolrunningtapwaterfor5minutes.

        14.PlaceslidesinGills#1hematoxylinfor90seconds.

        15.Placeslidesincoolrunningtapwaterfor5minutes.

        16.DehydrateslidesthroughbathsofA)70%ethylalcohol,B)80%ethylalcohol,C)90%ethylalcohol,D)95%ethylalcoholx2,E)100%ethylalcoholx2,F)xylene/100%ethanol1:1(orHemo-D/100%ethanol1:1)x2,G)xyleneorHemo-Dx2(2minuteseachbath.

        17.Mountslidesusingmountingmedia.

        FROZENSECTIONS(cutat8microns):Pullsectionsfromfreezer(orcutfresh),airdryatroomtemperature(orunderafanfor15minutes),fixedin4oCacetonefor4minutes,airdryfor30minutesatroomtemperature(orunderafanfor15minutes),thenfollowaboveprocedurebeginningat#5.

        SMEARS:Methanolfixfor30minutesat4oC,thenfollowaboveprocedurebeginningat#5.

        CYTOSPINS:CelllinesandothercellpopulationspreparedbyresUSPending,ineachwell,~40,000cellsusing500mL1%NormalGoatSerumperwell.Centrifugefor6minutesat1200RPM.Airdryfor30minutesatroomtemperature(orunderafanfor15minutes).Iftumorcellsarebeingscreened,fixinRTacetonefor2-3minutes.Ifperipheralbloodprepsarebeingscreened,fixin100%methanolfor20minutes.Airdrytheslidesfor30minutesatroomtemperature(or15minutesunderafan)andbeginat#5above.

        NOTE:WhenstainingusingthePCNAantibodiesastheprimaryDONOTuseNormalGoatSerumoranyotherserumblockingstep.
        BIOLOGicalInformation
        ImmunogenPurifiedPCNA
        Clone131-11912
        HostMouse
        SpecificitySpecificforPCNAa36kDaproliferationassociatedantigen.
        IsotypeIgM
        SpeciesReactivity
        • Bovine
        • Canine
        • Human
        • Mouse
        • Rat
        • Rabbit
        • Fish
        • Vertebrates
        AntibodyTypeMonoclonalAntibody
        EntrezGeneNumber
        EntrezGeneSummaryTheproteinencodedbythisgeneisfoundinthenucleusandisacofactorofDNApolymerasedelta.TheencodedproteinactsasahomotrimerandhelpsincreasetheprocessivityofleADIngstrandsynthesisduringDNAreplication.InresponsetoDNAdamage,thisproteinisubiquitinatedandisinvolvedintheRAD6-dependentDNArepairpathway.Twotranscriptvariantsencodingthesameproteinhavebeenfoundforthisgene.Pseudogenesofthisgenehavebeendescribedonchromosome4andontheXchromosome.
        GeneSymbol
        • PCNA
        • Cyclin
        • cyclin
        • MGC8367
        UniProtNumber
        UniProtSummaryFUNCTION:SwissProt:P12004#ThisproteinisanauxiliaryproteinofDNApolymerasedeltaandisinvolvedinthecontrolofeukaryoticDNAreplicationbyincreasingthepolymerase"sprocessibilityduringelongationoftheleadingstrand.
        SIZE:261aminoacids;28769Da
        SUBUNIT:Homotrimer.InteractswithKCTD10(Bysimilarity).Formsacomplexwithactivator1heteropentamerinthepresenceofATP.InteractswithPOLH,POLK,DNMT1,ERCC5/XPG,FEN1,CDC6,APEX2andPOLDIP2.InteractswithEXO1andSHPRH.
        SUBCELLULARLOCATION:Nucleus.
        PTM:Uponmethylmethanesulfonate-inducedDNAdamage,mono-ubiquitinatedbytheUBE2B-RAD18complexonLys-164.Thisinducesnon-canonicalpoly-ubiquitinationonLys-164through"Lys-63"linkageofubiquitinmoietiesbytheE2complexUBE2N-UBE2V2andtheE3ligaseSHPRH,whihisrequiredforDNArepair.
        DISEASE:SwissProt:P12004#Antibodiesarepresentinserafrompatientswithsystemiclupuserythematosus.
        SIMILARITY:SwissProt:P12004##BelongstothePCNAfamily.
        PhysicochemicalInformation
        Dimensions
        MaterialsInformation
        MaterialsInformation
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