ReviewProteinblotting:areviewBijiT.Kuriena,*,R.HalScofielda,b,caArthritisandImmunologyProgram,OklahomaMedicalResearchFoundation,825NE13thStreet,OklahomaCity,OK73104,USAbDepartmentofMedicine,UniversityofOklahomaHealthSciencesCenter,OklahomaCity,OK73104,USAcDepartmentofVeteransAffairsMedicalCenter,OklahomaCity,OK73104,USAReceived26August2002;receivedinrevisedform15November2002;accepted22November2002Keywords:Westernblotting;SDS-PAGE;Nitrocellulose1.IntroductionTheutilityofthehighresolvingpowerofgelelectrophoresiswaslimitedinpurpose,owingtothefactthattheseparatedproteinsinthegelmatrixweredifficulttoaccesswithmolecularprobes,untiltheadventofaprocedurethatallowedtransferofproteinsfromthegeltoanadsorbentmembrane(Towbinetal.,1979).ProteinblottingevolvedasanoffshootofDNA(Southern)blotting(Southern,1975)andRNA(North-ern)blotting(AlwineandKemp,1977).Asanefforttoretainthe‘geographic’namingtraditioninitiatedbySouthern(1975),Burnette(1981)coinedtheterm‘Westernblotting’todescribeaslightlymodifiedprocedurefromthatofTowbinetal.(1979).Westerntransferwithsubsequentimmunodetectionisapower-fultooltodetectandcharacterizeamultitudeofproteins,especiallythoseproteinsthatareoflowabundance.Itinvolvesthetransferofproteinssepa-ratedonsodiumdodecylsulfatepolyacrylamidegelelectrophoresis(SDS-PAGE)(Laemmli,1970)toasolidsupport.Theblottedproteinsformanexactreplicaofthegelandhaveprovedtobethestartingstepforavarietyofexperiments.Thesubsequentemploymentofantibodyprobesdirectedagainstthenitrocelluloseboundproteinshasrevolutionizedthefieldofimmunology.Sinceitsinception,proteinblot-tinghasbeenevolvingconstantlyandnowthescien-tificcommunityisconfrontedwithaplethoraofwaysandmeansoftransferringproteins.1.1.EfficiencyofproteinblottingTwomajorfactorsaffecttheefficiencyofproteinblotting:(1)theelutionefficiencyofaproteinoutofagelmatrixand(2)theefficiencyofbindingbythemembrane.1.1.1.TheelutionefficiencyofaproteinoutofagelmatrixAllblottingtechniquesrequireasuccessfultransferofproteinsfromthegeltoasolidmemb...
