DescriptionCatalogueNumberMAB377BrandFamilyChemicon®TradeNameChemiconDescriptionAnti-NeuNAntibody,cloneA60AlternateNamesNeuron-SpecificNuclearProteinNeuna60A60BackgroundInformationNeuNantibody(NEUronalNuclei;cloneA60)specificallyrecognizestheDNA-binding,neuron-specificproteinNeuN,whichispresentinmostCNSandPNSneuronalcelltypesofallvertebratestested.NeuNproteindistributionsareapparentlyrestrictedtoneuronalnuclei,perikaryaandsomeproximalneuronalprocessesinbothfetalandadultbrainalthough,someneuronsfailtoberecognizedbyNeuNatallages:INLretinalcells,Cajal-Retziuscells,Purkinjecells,inferiorolivaryanddentatenucleusneurons,andsympatheticganglioncellsareexamples(Mullenetal.,1992;Wolfetal.,1996).ImmunohistochemicallydetectableNeuNproteinfirstappearsatdevelopmentaltimepointsthatcorrespondwiththewithdrawaloftheneuronfromthecellcycleand/orwiththeinitiationofterminaldifferentiationoftheneuron(Mullenetal.,1992).ImmunoreactivityappearsaroundE9.5inthemouseneuraltubeandisextensivethroughoutthedevelopingnervoussystembyE12.5.Strongnuclearstainingsuggestsanuclearregulatoryproteinfunction;however,noevidencecurrentlyexistsastowhethertheNeuNproteinantigenhasafunctioninthedistalcytoplasmorwhetheritismerelysynthesizedtherebeforebeingtransportedbackintothenucleus.Nodifferencebetweenproteinisolatedfrompurifiednucleiandwholebrainextractonimmunoblotshasbeenfound(Mullenetal.,1992).ProductInformationFormatPurifiedControlPositivecontrol-BrainTissue.Negativecontrol-AnynonneuronaltissueegFibroblastsPresentationPurifiedmouseimmunoglobulinIgG1liquidinbuffercontaining0.02Mphosphatebuffer,0.25MNaCl,pH7.6with0.1%sodiumazide.StorageandShippingInformationStorageConditionsStablefor6monthsat2-8ºCfromdateofreceipt.ApplicationsApplicationAnti-NeuNAntibody,cloneA60detectslevelofNeuNandhasbeenpublishedandvalidatedforuseinFC,IC,IF,IH,IH(P),IPandWB.KeyApplicationsFlowCytometryImmunocytochemistryImmunofluorescenceImmunohistochemistryImmunohistochemistry(Paraffin)ImmunoprecipitationWesternBlottingApplicationNotesWesternBlotAnalysis:Apreviouslotofthisantibodyrecognized2-3bandsinthe46-48kDarangeandpossIBLyanotherbandatapproximately66kDa.Immunocytochemistry:1:10-1:100dilutionfromapreviouslotwasused.Neuronsincultureshouldbepermeablizedwith0.1%tritonX-100.Allprimaryantibodydilutionsshouldbeperformedwithsimplesolutionscontainingonlybufferandprimaryantibodywithoutexcessproteinblocksordetergents.Immunohistochemistry:1:100-1:1,000.Theantibodyworksbestonpolyesterwaxembeddedtissuebutalsoworksonparaffinembeddedtissueatalowerworkingdilution.Theantibodyworkswellwithformaldehyde-basedfixatives.Citricacidandmicrowavepretreatmenthasbeenusedsuccessfully(Sarnat,1998).Immunohistochemistry(paraffin)Analysis:ApreviouslotwasusedforIH(P).Optimalworkingdilutionsmustbedeterminedbyenduser.BIOLOGicalInformationImmunogenPurifiedcellnucleifrommousebrainCloneA60ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.HostMouseSpecificityMillipore"sexclusivemonoclonalantibodytovertebrateneuron-specificnuclearproteincalledNeuN(orNeuronalNuclei)reactswithmostneuronalcelltypesthroughoutthenervoussystemofmiceincludingcerebellum,cerebralcortex,hippocampus,thalamus,spinalcordandneuronsintheperipheralnervoussystemincludingdorsalrootganglia,sympatheticchaingangliaandentericganglia.Developmentally,immunoreactivityisfirstobservedshortlyafterneuronshavebecomepostmitotic,nostaininghasbeenobservedinproliferativezones.Theimmunohistochemicalstainingisprimarilylocalizedinthenucleusoftheneuronswithlighterstaininginthecytoplasm.ThefewcelltypesnotreactivewithMAB377includePurkinje,mitralandphotoreceptorcells.TheantibodyisanexcellentMarkerforneuronsinprimaryculturesandinretinoicacid-stimulatedP19cells.Itisalsousefulforidentifyingneuronsintransplants.IsotypeIgG1SpeciesReactivityAvianChickenFerretHumanMousePigRatSalamanderAntibodyTypeMonoclonalAntibodyGeneSymbolNeuNA58PurificationMethodProteinAPurfiedMolecularWeight46/48kDaPhysicochemicalInformationDimensionsMaterialsInformationMaterialsInformation