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Millipore/MAB4072 | Anti-BrdU Antibody, clone 131-14871/MAB4072/100 µg
  • Millipore/MAB4072 | Anti-BrdU Antibody, clone 131-14871/MAB4072/100 µg

Millipore/MAB4072 | Anti-BrdU Antibody, clone 131-14871/MAB4072/100 µg

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貨號(hào): MAB4072
品牌: Millipore
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    • Description
      CatalogueNumberMAB4072
      BrandFamilyChemicon®
      TradeName
      • Chemicon
      DescriptionAnti-BrdUAntibody,clone131-14871
      AlternateNames
      • BrdU
      BackgroundInformationThethymidineanalog5-bromo-2′-deoxyuridine(BrdU)isacommonreagentusedforbothcellproliferationassaysandforthedetectionofapoptoticcells.BrdUisreADIlyincorporatedintonewlysynthesizedDNA,labelingcellsthathaveprogressedthroughtheS-phaseofthecellcycleandtherebyservingasaMarkerforproliferatingcells.
      ProductInformation
      FormatPurified
      Control
      • Brdutreatedcells
      PresentationLiquidin10mMPhosphatebuffer,pH7.4containing150mMNaCland0.1%sodiumazide.
      StorageandShippingInformation
      StorageConditionsMaintainfor2yearsat-20°Cfromdateofshipment.Aliquottoavoidrepeatedfreezingandthawing.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.
      Applications
      ApplicationAnti-BrdUAntibody,clone131-14871isaMouseMonoclonalAntibodyfordetectionofBromodeoxyuridinealsoknownasBrdU&hasbeenvalidatedinFC,IHC,IHC(P).Thisbromodeoxyuridineantibodyisexpecttoreactinallspecies.
      KeyApplications
      • FlowCytometry
      • Immunohistochemistry
      • Immunohistochemistry(Paraffin)
      ApplicationNotesImmunohistochemistryondeparaffinizedsectionsofkidney,duodenum,andlivertissue:1:1000-1:1250.

      Flowcytometryon100μLof106cells:1:5000.

      Optimaldilutionsmustbedeterminedbytheenduser.

      IMMUNOHISTOCHEMICALPROCEDURE

      FORPARAFFINEMBEDDEDTISSUESECTIONS

      STAININGPROCEDURE:

      1.Deparaffinizesectionsinxylene3x5minutes.ResinembeddedorGMA(Glycolmethacrylate,2-hydroxyethylmethacrylate)treatedsectionsitisunnecessarytodeparaffinizeasthereisnoparaffin.

      2.Placeslidesingradedalcohols(100,95,90%)towaterat2minuteintervals,andairdry.(OmitforGMAsections)

      3.CirclesectionswithPAPpenordiamondpentoidentifyanddrythoroughly.

      4.ForGMAorresinsections,placeslidesin0.5%Tween/PBS(pH7.6)for29minutes.

      5.Incubateinprewarmed(40°C)1NHClfor1hour.

      6.Wash3x3minuteswithPBS(pH7.6).Slidesmaybeheldatthispointandtheprocedurecontinuedthefollowingday.

      7.Incubatein1XTrypsinsolution(0.02-0.05%w/v,prewarmedto40°C)for20minutesatroomtemperatureforNBF(normalbufferedformalin)fixedtissueor10minutesforCarnoy"sfixedtissue.

      8.Wash3x5minuteswithPBS(pH7.6).

      9.Incubatesectionsin1%H2O2(10mL30%H2O2in290mLmethanol)for20minutes,orGMAsectionsin3%H2O2(10mL30%H2O2in90mLwater)for3minutes.

      10.Wash2x2minuteswithPBS(pH7.6).

      11.UsingtheVectorABCkit,add3dropsofnormalhorseserumto10mLofPBS.Usethissolutiontoblocktheslidesfor20minutesatroomtemperature.

      12.BlottheslidesofexcessnormalhorseserumandincubatewithMAB4072dilutedinPBS/BSA/Tween20for1houratroomtemperature.

      13.Wash3x3minuteswithPBS(pH7.6).

      14.Add3dropsofnormalhorseserumto10mLofPBSandthenadd1dropofbiotinylatedantibodystock.Incubatetheslideswiththisdilutedsecondantibodyfor30minutesatroomtemperature.

      15.PreparetheVectastainABCReagentbyadding2dropsofreagentAto5mLofPBS.Thenadd2dropsofReagentBtothesamemixingbottle.Allowtositforabout30minutespriortouse.

      16.Wash3x3minuteswithPBS(pH7.6).

      17.IncubatetheslideswithABCReagentfor30minutesatroomtemperature.

      18.Wash3x3minuteswithPBS(pH7.6).

      19.Preparethesubstratebymixinganequalvolumeof0.02%H2O2(madeindistilledwaterfroma30%stocksolution)and0.1%DABmadein0.1MTrisbuffer,pH7.2.TheH2O2shouldbefreshlypreparedfromconcentratedstock.BecausemanyperoxidesubstratesareunstableinthepresenceofH2O2orwhenexposedtolight,thesubstrateshouldbepreparedjustpriortouse.SinceDABisasUSPectedcarcinogen,careshouldbetakeninhandlinganddisposingofallperoxidasesubstrates.

      20.StaintheslideswithDABsubstratefor2-5minutes.

      21.Wash2x2minuteswithdistilledwater.

      22.Counterstainwithhemotoxylin,dehydratethroughxylene,andmountthecoverslip.
      BIOLOGicalInformation
      Clone131-14871
      ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
      HostMouse
      SpecificityRecognizesBrdU(Bromodeoxyuridine).
      IsotypeIgG1
      SpeciesReactivity
      • All
      AntibodyTypeMonoclonalAntibody
      PurificationMethodProteinAPurfied
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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