ProductDescriptionSize:100μlAbType:pReactivity:AllImmunogen:NitratedKLHHost:rabbitProductDescriptionTherabbit-derivedanti-3NTantibodyispurifiedbynitratedtyrosineagaroseaffinitychromatography.Itspecificallyrecognizesproteinswithtyrosinenitratedresiduesbutnotthetyrosinephosphorylatedornon-modifiedtyrosineresidues.BesidescommonapplicationsinWesternblottingandimmunofluorescence,theantibodyhasbeenwellutilizedforproteomicscreeningtotyrosinenitratedsubstratesfromprotease-digestedproteinsbothinprokaryoticandeukaryoticcells.Source/PurificationThisproductisproducedbyimmunizingrabbitswithnitratedKLH.TheantiserumwasimmunoaffinitypurifiedbyproteinAandfollowedwithnitrotyrosinecontainingpeptides.SpecificityAlltyrosinenitratedproteinsexpectedRecommendedApplicationsELISA,WB,IP*WB=Westernblotting; IP=ImmunopreciptationRecommendedantibodydilutionWB:1:1000ImportantForWesternblotting,incubatemembranewithdilutedantibodyin5%BSA,1xTBS,0.1%Tween-20at4℃withgentleshaking,overnight.Useatanassaydependentconcentration.Optimaldilutions/concentrationsshouldbedeterminedbytheenduser.Notyettestedinotherapplications.Dotblottinganalysis onindicatedamountofpeptidesusing3-nitrotyrosinerabbitpAb.Thelistofpeptidesisincludedinthetableontheright.Westernblotanalysis on30μgofextractsfromHeLaandMEFwholecelllysates,untreatedortreatedwithperoxynitrite(3mM,30mins)orpervanadate(3mM,30mins),usinganti-3-nitrotyrosineAntibody(FigureB)oranti-phospho-TyrosinerabbitpAb(PTM-702)(FigureC).ScientificDescriptionNitrotyrosineisaproductoftyrosinenitrationmediatedbyreactivenitrogenspeciessuchasperoxynitriteanionandnitrogendioxide.ItisverystableandsoissuitableforanalysisinBIOLOGicalsamplesuchasplasma,lungaspirants-BALF(Bronchoalveolarliningfluid)andurine.IncreasednitrotyrosinelevelshavebeenassociatedwithseveralphysiologicalandpathologicalconditionsandisconsideredaMarkerofNO-dependent,reactivenitrogenspecies-inducednitrativestress.ResearchshowsthatnitrotyrosinelevelscanbereducedbyN-acetylcysteinewhichisaprecursortoglutathione,oneofthebody’sprimaryendogenousantioxidants.Storage&StABIlityTheantibodyisdeliveredinlyophilizedpowder.Uponreceipt,pleasecentrifugetheantibodyat12,000xgfor20seconds,andthenreconstituteitwiththeantibodystabilizerprovided.ThestabilizercontainsPBS,50%glycerol,and0.01%sodiumazide.Pleasestoretheantibodyat-20°C.Avoidrepeatedfreeze/thawcycles.Antibodyisstablefor12monthsfromdateofreconstitution.Equilibratetheantibodytoroomtemperaturefor2minutesandmixbygentlypipettingbeforeuse.**Researchpurposesonly.Theproductisnotintendedtobeusedfortherapeuticordiagnosticpurposesinhumansoranimals.