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Millipore/MAB351R | Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6/MAB351R/100 µg
  • Millipore/MAB351R | Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6/MAB351R/100 µg

Millipore/MAB351R | Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6/MAB351R/100 µg

價格: ￥4812.00 市場價: 8020.00
貨號: MAB351R
品牌: Millipore
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    • Description
      CatalogueNumberMAB351R
      BrandFamilyChemicon®
      TradeName
      • Chemicon
      DescriptionAnti-GlutamateDecarboxylaseAntibody,65kDaisoform,cloneGAD-6
      AlternateNames
      • GAD65
      BackgroundInformationGutamicaciddecarboxylase(GAD;E.C.4.1.1.15)istheenzymeresponsIBLefortheconversionofglutamicacidtogamma-aminobutyricacid(GABA),themajorinhibitorytransmitterinhigherbrainregions,andputativeparacrinehormoneinpancreaticislets.TwomolecularformsofGAD(65kDaand67kDa,64%aaidentitybetweenforms)arehighlyconservedandbothformsareexpressedintheCNS,pancreaticisletcells,testis,oviductandovary.Theisoformsareregionallydistributedcytoplasmicallyinthebrainsofratsandmice(Sheikh,S.etal.1999).GAD65isanampiphilic,membrane-anchoredprotein(585aa),encodedonhumanchromosome10,andisresponsibleforvesicularGABAproduction.GAD67iscytoplasmic(594aa.),encodedonchromosome2,andseemstoberesponsibleforsignificantcytoplasmicGABAproduction.GADexpressionchangesduringneuraldevelopmentinratspinalcord.GAD65isexpressedtransientlyincommissuralaxonsaroundE13butisdownregulatedthenextdaywhileGAD67expressionincreasesmostlyinthesomataofthoseneurons(Phelps,P.etal.1999).Inmatureratpancreas,GAD65andGAD67appeartobedifferentiallylocalized,GAD65primarilyininsulin-containingbetacellsandGAD67inglucagon-containing(A)cells(Li,L.etal.1995).GAD67expressionseemstobeparticularlyplasticandcanchangeinresponsetoexperimentalmanipulation(forexampleneuronalstimulationortransection)ordiseaseprogressionandemergentdisorderslikeschizophrenia(Volketal.,2000).ColocalizationofthetwoGADisoformsalsoshowschangesinGAD65/GAD67distributionscorrelatedwithcertaindiseasestatessuchasIDDMandSMS.
      ProductInformation
      FormatPurified
      Control
      • Braintissue
      PresentationLyophilized.Dissolvecontentsofvialin100μLofsterile,distilledwater.Thisresultsinafinalantibodyconcentrationof1mg/mlin10mMpotassiumphosphate,70mMsodiumchloride,pH7.4containingnopreservatives.
      StorageandShippingInformation
      StorageConditionsMaintainfor1yearat-20°Cfromdateofshipment.Aliquottoavoidrepeatedfreezingandthawing.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.
      Applications
      ApplicationAnti-GlutamateDecarboxylaseAntibody,65kDaisoform,cloneGAD-6isanantibodyagainstGlutamateDecarboxylaseforuseinIH&WB.
      KeyApplications
      • Immunohistochemistry
      • WesternBlotting
      ApplicationNotesImmunohistochemistry:(<=1=""μg/ml)=""optimal=""working=""dilutions=""must=""be=""determined=""by=""end=""user.="">



      ImmunohystochemicalStainingProcedures

      ThefollowingprocedurewasdevelopedtolocalizeGADinratbrainsectionsofcerebellum.Performallstepsatroomtemperatureunlessotherwiseindicated.Wherenormalserumisindicated,usenormalserumfromthesamespeciesasthesourceofthesecondaryantibody.Thisprocedurerepresentssuggestedguidelinesfortheuseofanti-GAD.Fixationregimen,antibodyconcentrations,andincubationconditionsforagivenexperimentalsystemshouldbedeterminedempirically.

      1.Perfuseratswith100mMphosphatebuffer,pH7.4,containing1%paraformaldehyde,0.34%L-lysine,and0.05%sodiumm-periodate(1%PLP).

      2.Postfixbrainsin1%PLPfor1-2hours.Longerfixationtimesmayreducelabelingintensity.

      3.Transferbrainsto100mMphosphatebuffercontaining30%sucrose,andgentlyagitateonashakerplatformat+4°Cfor48-60hours.

      4.Usingaslidingmicrotome,cut30mmsectionsoffrozencerebellum.Asthesectionsarecut,collecttheminavialofcold100mMphosphatebuffer.

      5.Incubatesectionsinphosphate-bufferedsaline(PBS)containing1.5%normalserumand0.2%TritonX-100for30minutes.

      6.Onashakerplatform,incubatesectionswithanti-GAD(dilutedinPBScontaining1.5%normalserumand0.2%TritonX-100toafinalantibodyconcentrationof1mg/ml)for12-36hoursat+4°C.

      7.Onashakerplatform,rinsesectionseighttimes,10-15minutesperrinse,inPBS.

      8.Detectwithastandardsecondaryantibodydetectionsystem(Hsuetal.,1981;Falini&Taylor,1983;Harlow&Lane,1988;Taylor,1978).

      9.Mountsections,dehydrate,andapplycoverslips.
      BIOLOGicalInformation
      ImmunogenPurifiedratbrainGAD.
      CloneGAD-6
      ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
      HostMouse
      SpecificityRecognizesthelowermolecularweightisoformofthetwoGADisoformsidentifiedinbrain(Gottlieb,etal.,1986;Chang&Gottlieb,1988).Thismonclonalantibodycanbeusedforimmunohistochemicallocalizationinbrainorpancreas.Anti-GADhasalsobeenusedtolabelpurifiedGADonWesternblots(Chang&Gottlieb,1988).
      IsotypeIgG2a
      SpeciesReactivity
      • Human
      • Rat
      AntibodyTypeMonoclonalAntibody
      EntrezGeneNumber
      EntrezGeneSummaryThisgeneencodesoneofseveralformsofglutamicaciddecarboxylase,identifiedasamajorautoantigenininsulin-dependentdiabetes.Theenzymeencodedisresponsibleforcatalyzingtheproductionofgamma-aminobutyricacidfromL-glutamicacid.ApathogenicroleforthisenzymehasbeenidentifiedinthehumanpancreassinceithasbeenidentifiedasanautoantibodyandanautoreactiveTcelltargetininsulin-dependentdiabetes.Thisgenemayalsoplayaroleinthestiffmansyndrome.
      GeneSymbol
      • GAD2
      • MGC161607
      • GAD65
      • MGC161605
      • GAD-65
      • EC4.1.1.15
      PurificationMethodAmmoniumsulfateprecipitationandDEAE-cellulosechromatography
      UniProtNumber
      UniProtSummaryFUNCTION:SwissProt:Q05329#CatalyzestheproductionofGABA.
      COFACTOR:Pyridoxalphosphate.
      SIZE:585aminoacids;65411Da
      SUBUNIT:Homodimer(Bysimilarity).
      SUBCELLULARLOCATION:Cytoplasm,cytosol.Cytoplasmicvesicle.Celljunction,synapse,presynapticcellmembrane;Lipid-anchor.Golgiapparatusmembrane;Peripheralmembraneprotein;Cytoplasmicside.Note=Associatedtocytoplasmicvesicles.Inneurons,cytosolicleafletofGolgimembranesandpresynapticclusters.
      PTM:Phosphorylated;whichdoesnotaffectkineticparametersorsubcellularlocation.&Palmitoylated;whichisrequiredforpresynapticclustering.
      SIMILARITY:SwissProt:Q05329##BelongstothegroupIIdecarboxylasefamily.
      MolecularWeight65kDa
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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