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Millipore/MAB305 | Anti-Choline Acetyltransferase Antibody, clone 1E6/MAB305/100 µL
  • Millipore/MAB305 | Anti-Choline Acetyltransferase Antibody, clone 1E6/MAB305/100 µL

Millipore/MAB305 | Anti-Choline Acetyltransferase Antibody, clone 1E6/MAB305/100 µL

價(jià)格: ￥3072.00 市場(chǎng)價(jià): 5120.00
貨號(hào): MAB305
品牌: Millipore
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    • Description
      CatalogueNumberMAB305
      BrandFamilyChemicon®
      TradeName
      • Chemicon
      DescriptionAnti-CholineAcetyltransferaseAntibody,clone1E6
      AlternateNames
      • ChAT
      • CholineAcetylase
      • CHOACTase
      ProductInformation
      FormatAscites
      Control
      • Braintissue
      PresentationAscitesfluidcontainingnopreservatives.
      StorageandShippingInformation
      StorageConditionsMaintainfor1yearat-20°Cfromdateofshipment.Aliquottoavoidrepeatedfreezingandthawing.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.
      Applications
      ApplicationDetectCholineAcetyltransferaseusingthisAnti-CholineAcetyltransferaseAntibody,clone1E6validatedforuseinIH.
      KeyApplications
      • Immunohistochemistry
      ApplicationNotesImmunohistochemistry:1:100-1:250.Seeimmunohistochmistryprocedurebelow.

      Optimalworkingdilutionsmustbedeterminedbytheenduser.

      IMMUNOHISTOCHEMISTRYPROCEDURE(PAPTECHNIQUE)FORMAB305,MONOCLONALANTIBODYTOCHOLINEACETYLTRANSFERASE

      I)Perfusion&SectioningProcedure

      1.Perfusethroughtheheartwithafixativesolutioncontaining4%paraformaldehydein0.12Mphosphatebuffer(pH7.3)forlightmicroscopy(LM),andadditionally,0.1%gluteraldehydeand.002%CaCl2forelectronmicroscopy(EM).

      2.Removebrainandpostfix2-18hoursat4°Cin4%paraformaldehydein0.12Mphosphatebuffer.

      3.Afterbrainisblockedforsectioning,washinseveralchangesofbufferfor2-3hours.

      4.SpecimensforEMaresectionedonaVibratome(50μm)andrinsedinbuffer,thoseforLMshouldbecryoprotectedin30%sucroseinbuffer.

      5.Afterfreezingwithdryice,30-40μmthicksectionsofLMspecimensarecutonacryostat.

      6.Sectionsarerinsed,andthenstoredinphosphatebuffercontaining0.1%sodiumazide.

      II)StainingProcedure

      Tissueisprocessedasfreely-floatingsectionsincontinuouslyagitatedsolutions.Allincubationsareperformedatroomtemperatureunlessotherwisestated.

      1.a.ForlocalizingChAT-positivesomataanddendrites:

      Sectionsarewashedin0.1MTris-bufferedsaline(TBS;containing1.4%NaCl,pH7.3)only.Nodetergentorenzymepretreatmentisused.

      b.ForlocalizingChAT-positiveterminal-likestructures:

      IncubatesectionsinTBS(pH8.1)for5minutesat37°C.TransfersectionstoTBS(pH8.1)containingpronase(1.2μg/mL)for11/2-2minutesat37°C,followedbyseveralicecoldbufferwashesforatotalof5minutes.Theconcentrationofpronaseandincubationtimeofthedigestionshouldbeevaluatedforeachregionexamined.

      c.ForlocalizingChATimmunoreactivityandsubsequentlycounterstainingthesections:

      IncubationinTBScontaining0.1%-0.8%TritonX-100for15minutesmayincreasethetissuepenetrationoftheImmunoReagents,butitalsoraisesthebackgroundstaining.

      2.Incubatesectionsinnormalgoatserum(3-5%)foronehour.Theworkingsolutionsofallantiserashouldalsocontainsimilarlydilutednormalgoatserum.

      3.Incubateinanti-ChATmonoclonalantibodysolution(Suggestedworkingdilution1:250,finalworkingdilutionmustbedeterminedbyenduser)for2hoursatroomtemperatureandthenforanadditional6-18hoursat4°C.

      4.Incubatewithsecondantibody(i.e.Goatanti-MouseIgG,Cat.No.:AP124,dilution1:50-100)for1-2hours.

      5.IncubatewithdilutedPAPcomplex(i.e.MousePAP,CatNo.:PAP14,conc.25-50μg/mL)foronehour.

      6.Afterrinsinginbuffer,thesecondantibodyandPAPstepsarerepeatedfor40minutesto1houreachinordertoamplifystainingintensity,particularlyofsmallChAT-containingstructures.

      7.Reactfor15minuteswith0.06%3,3"-diaminobenzidine×4HCl(DAB;dilutedinphosphatebufferedsaline,pH7.3)and0.006%H2O2.

      8.SpecimensforroutineLMarepostfixedfor1minutesin0.005%OsO4(osmiumtetraoxide),andthenmounted,dehydratedandcoverslipped.SelectedregionsblockedforEMarepostfixedin2%OsO4for1hour,enblocstainedwithuranylacetate,andflat-embeddedinEpon-Aralditeresin.
      BIOLOGicalInformation
      ImmunogenCholineacetyltransferasepurifiedfromratbrain.
      Clone1E6
      ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
      HostMouse
      SpecificityRecognizescholinergicneuronsinthebrainandspinalcord(CNS).
      IsotypeIgG1
      SpeciesReactivity
      • Human
      • Monkey
      • Rat
      AntibodyTypeMonoclonalAntibody
      EntrezGeneNumber
      EntrezGeneSummaryCholinergicsystemsareimplicatedinnumerousneurologicfunctions.AlterationinsomecholinergicneuronsmayaccountforthedisturbancesofAlzheimerdisease.Theproteinencodedbythisgenesynthesizestheneurotransmitteracetylcholine.Alternativesplicevariantshavebeenfoundthatcontainalternative5"untranslatedexons.Threeofthefourdescribedsplicevariantsencodeidentical69kDaproteinswhileonevariantencodesboththe69kDaandalarger82kDaprotein.
      GeneSymbol
      • CHAT
      • ChAT
      • CMS1A2
      • CHOACTase
      • CMS1A
      • EC2.3.1.6
      PurificationMethodUnpurified
      UniProtNumber
      UniProtSummaryFUNCTION:SwissProt:P28329#CatalyzesthereversIBLesynthesisofacetylcholine(ACh)fromacetylCoAandcholineatcholinergicsynapses.
      SIZE:748aminoacids;82568Da
      DISEASE:SwissProt:P28329#DefectsinCHATarethecauseoffamilialinfantilemyastheniagravis2(FIMG2)[MIM:254210,254200];alsoknownasCMS-EA.FIMG2patientshavemyasthenicsymptomssincebirthorearlyinfancy,negativetestsforanti-AChRantibodies,andabruptepisodiccriseswithincreasedweakness,bulbarparalysis,andapneaprecipitatedbyundueexertion,fever,orexcitement.Inheritanceisautosomalrecessive.
      SIMILARITY:SwissProt:P28329##Belongstothecarnitine/cholineacetyltransferasefamily.
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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