TheDetectX^®DNADamageImmunoassayKitisdesignedtoquantitativelymeasureDNAandRNAoxidizedguanosinespecies.Theassaydetectsallthreeoxidizedguaninespecies;8-hydroxy-2’-deoxyguanosinefromDNA,8-hydroxyguanosinefromRNAand8-hydroxyguaninefromDNAorRNA.Thesespeciesmaybepresentinserum,plasma,saliva,urine,driedfecalsamples,andtissueculturemediasamples.

An8-hydroxy-2’-deoxyguanosine(8-OHdG)stocksolutionisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.StandardsordilutedsamplesarePipettedintoaclearmicrotiterplatecoatedwithanantibodytocapturerabbitantibodies.An8-hydroxyguanosineconjugateisaddedtothestandardsandsamplesinthewells.Thebindingreactionisinitiatedbytheadditionofaperoxidase-labeledmousemonoclonalantibodyto8-hydroxy-2’-deoxyguanosinetoeachwell.Aftera2hourincubationtheplateiswashedandsubstrateadded.Thesubstratereactswiththeperoxidaselabeledantibodythathasreactedwiththeboundconjugate.Afterashortincubation,thereactionisstoppedandtheintensityofthegeneratedcolorisdetectedinamicrotiterplatereadercapableofmeasuring450nmwavelength.Theconcentrationofthe8-hydroxy-2’-deoxyguanosineinthesampleiscalculated,aftermakingsuitablecorrectionforthedilutionofthesample,usingsoftwareavailablewithmostplatereaders.