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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(jià)格: ¥3576.00 市場(chǎng)價(jià): 5960.00

貨號(hào): 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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