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alpha-internexinisa~66kDaClassIVintermediatefilamentoriginallydiscoveredasitcopurifieswithotherneurofilamentsubunits.Itisrelatedtobutdistinctfromthebetterknownneurofilamenttripletproteins,NF-L,NF-MandNF-H,havingsimilarproteinsequencemotifsandasimilarintronorganization.Itisexpressedinlargeamountsearlyinneuronaldevelopment,butisdownregulatedinmanyneuronsasdevelopmentprocedes.Manyclassesofmatureneuronscontainalpha-internexininadditiontoNF-L,NF-ManbdNF-H.Insomematureneuronsalpha-internexinistheonlyneurofilamentsubunitexpressed.Antibodiestointernexinarethereforeuniqueprobestostudyandclassifyneuronaltypesandfollowtheirprocessesinsectionsandintissueculture.Inadditiontheveryearlydevelopmentalexpressionofalpha-internexinmeansitspresenceisanearlyandconvenientdiagnosticfeatureofneuronalprogenitorscellsandothercellcommittedtotheneuronallineage.
Image:MixedneurongliaculturesstainwithchickenNeurofilamentalpha-internexin/NF66(red)andcounterstainedwithCoronin1a,(green)whichisanexcellentMarkerofmicrogliaandlymphocytes.Thechickeninternexinantibodyisanexcellentmarkerofneuronalprocessesinthesecultures.
Note:Theserumcanbedilutedto1:1,000andhigherforimmunofluorescencestainingand1:10,000andhigherforwesternblotting.OnwesternblotsuseabrainorspinalcordhomogenateandlookforamajorbandatSDS-PAGEapparentmolecularweightof66to68kDa,dependingonthespecies.Thehumanproteinrunsat~68kDa,whilerat,mouseandotherspeciesrunat~66kDa.Minorbandsat~150kDaareprobablycovalentdimers,andbandsat~50kDarepresenta-internexinbreakdownproducts.