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Millipore/AB5522 | Anti-Acetylcholine Antibody/AB5522/100 µL
  • Millipore/AB5522 | Anti-Acetylcholine Antibody/AB5522/100 µL

Millipore/AB5522 | Anti-Acetylcholine Antibody/AB5522/100 µL

價(jià)格: ￥4440.00 市場(chǎng)價(jià): 7400.00
貨號(hào): AB5522
品牌: Millipore
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    • Description
      CatalogueNumberAB5522
      BrandFamilyChemicon®
      TradeName
      • Chemicon
      DescriptionAnti-AcetylcholineAntibody
      ProductInformation
      FormatSerum
      PresentationRabbitantiserum.Liquidwith0.05%sodiumazide.
      StorageandShippingInformation
      StorageConditionsMaintainat-20°Cinundilutedaliquotsforupto6monthsafterdateofreceipt.Avoidrepeatedfreeze/thawcycles.



      Duringshipment,smallvolumesofantibodywilloccasionallybecomeentrappedinthesealoftheproductvial.Forantibodieswithvolumesof200μlorless,werecommendgentlytappingthevialonahardsurfaceorbrieflycentrifugingthevialinatabletopcentrifugetodislodgeanyliquidinthecontainer"scap.
      Applications
      ApplicationAnti-AcetylcholineAntibodyisanantibodyagainstAcetylcholineforuseinIH.
      KeyApplications
      • Immunohistochemistry
      ApplicationNotesImmunohistochemistry:1:250-1:2500byPAP(seesuggestedprotocolbelow).

      Optimalworkingdilutionsmustbedeterminedbytheenduser.

      PROTOCOLforAcetylcholineDetectionbyImmunohisto/cytochemistry.Exampleforaratbrain.

      1.SOLUTIONSTOBEPREPARED-Solutionmustbepreparedasneeded.

      SolutionA:0.5Mglutaraldehyde,0.1M2-nitrobenzylalcohol,10g/LsodiummetABIsulfite,0.1Mcacodylate,0.01Msodiumbromide,pH10.7.

      SometimesaprecipitatemayformwhilemakingsolutionA.Toavoidthis,firstprepareamixturewith1.0Mglutaraldehydeand0.2M2-nitrobenzylalcohol.Prepareasecondmixturewith20g/Lsodiummetabisulfiteand0.2Mcacodylate;addsodiumbromidetoobtainpH10.7.Thencombinethetwomixturesinequalpartswhilestirring.



      SolutionB:0.05MTris,10.0g/Lsodiummetabisulfite,pH7.4.(*)

      2.RATPERFUSION-TheratisanaesthetizedwithsodiumpentobarbitalorNembutalandperfusedintracardiallythroughtheaortausingapumpwithSolutionA(200-300mL/min).

      3.POSTFIXATION:2hoursin0.5Mglutaraldehydesolution,(pH7.5)withoutthe2-nitrobenzylalcohol,then4softwashesinsolutionB.

      4.TISSUESECTIONING:Vibratomeorcryostatsectionscanbeused.

      Thesectionsarewashed4timesinsolutionB,andincubatedfor1hourat37°CinsolutionBcontaining0.2%TritonX100,plus1%nonspecificserum.

      5.APPLICATIONOFACETYLCHOLINEANTIBODY:Useafinaldilutionof1:500-1:2,500inSolutionBcontaining0.2%TritonX100and1%non-specificserum.Incubate12sectionsper2mLdilutedantibodyovernight,+4°C.Thenwashthesectionsthreetimesfor10minuteseachinSolutionB.(Notethattheantibodymaybeusableatahigherdilution.Thisshouldbeexploredtominimizethepossibilityofhighbackground.Additionally,notethatachangeinthebufferingsystemasindicatedintheprotocolmaychangethebackgroundandantibodyrecognition).ThespecificreactionisthenrevealedbyPAPprocedure.

      6.SECONDANTIBODY:Incubatethesectionswitha1:50to1:200dilutionofgoatanti-rabbitinSolutionBcontaining1%non-specificserumforeither3hrsat20°Cor2hrat37°C.Thenwashthesections,3times,for10minuteseachwithSolutionB.

      7.PAP:Incubatethesectionswiththeappropriatedilutionofperoxidaseanti-peroxidase(forfreefloatingmethod)inSolutionBcontaining1%non-specificserumfor1-2hoursat37°C.Thenwashsections3timesfor10mineachinsolutionB.

      8.VISUALIZATION:Theantigen-antibodycomplexesarevisualizedusingDAB-4-HCl(25mg/100mL)(orotherchromogen)in0.05MTrisandfiltrated;0.05%hydrogenperoxideisadded.Incubatethesectionsfor10minutesatroomtemp.Stopthereactionbytransferringthesectionsto5mL0.05MTris.WashtissuewithsolutionDusing2,10minwashes.Mountsectionsonchrome-alumcoatedslides.Dryovernightat37°C.Rehydratesectionsusingconventionalhistologicalprocedures.Coverslipusingrapidmountingmedia.
      BIOLOGicalInformation
      ImmunogenAcetylcholine-Glutaricanhydride-PL
      HostRabbit
      SpecificityAcetylcholine



      Thecross-reactivitiesweredeterminedusingeitherELISAorRIAtechniques,atconcentration/unconjugatedorconjugatedaminoacidconcentrationathalfdisplacementwiththefollowingcompounds:



      CompoundCross-reactivity

      Choline-GA-PL1

      Phosphatidylcholine1/>10,000

      Choline1/>10,000

      Acetylcholine1/>10,000

      Choline-allylalcohol-G-BSA1/1.6



      Theantiserawasalsotestedforspecificityusingthefree-floatingPAPtechniqueonratcortex.



      Abbreviations:



      (GA)Glutaricanhydride

      (=)Non-reducedconjugate

      (PL)Polylysine

      (BSA)BovineSerumAlbumin
      SpeciesReactivity
      • Rat
      AntibodyTypePolyclonalAntibody
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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