TheEpiQuik™DNMT1Activity/InhibitorScreeningAssayCoreKitisaconvenientsetoftoolsthatallowstheexperimentertoselectivelyscreenDNAmethyltransferase1activity/inhibitorsinanELISA-likeformat.TheselectiveinhibitionofDNMT1mayleadtodemethylationandexpressionofsilencedtumorsuppressorgenes.Thus,theselectiveDNMT1inhibitorscouldbeanewadditiontocancertherapeuticagents.Thekitisready-to-useandprovidesalltheessentialcomponentsneededtocarryoutasuccessfulDNMT1activity/inhibitorscreeningexperimentwithouttheneedforrADIoactivityoranyspecialequipment.Thekithasthefollowingadvantages:

• Veryrapidprocedure,whichcanbecompletedwithin3hours.
• Safeandinnovativecolorimetricassaywithoutradioactivity,extraction,andchromatography.
• StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughputanalysis.
• Extremelysimple,reliable,andconsistentassayconditions.

Note:This"core"kitdoesnotcomewithenzymesincluded.

Principle&Procedure
TheEpiQuik™DNMT1Activity/InhibitorScreeningAssayKitisdesignedforscreeningDnmt1inhibitors.Inanassaywiththiskit,theuniquecytosine-rich DNAsubstrateisstablycoatedonthestripwells.ThesewellsarespecificallytreatedtohaveahighDNAabsorptionABIlity.ADNMT1enzyme(notincluded)transfersamethylgrouptocytosinefromAdomettomethylatetheDNAsubstrate.ThemethylatedDNAcanberecognizedwithananti-5-methylcytosineantibody.TheratiooramountofmethylatedDNA,whichisproportionaltoenzymeactivity,canthenbecolorimetricallyquantifiedthroughanELISA-likereaction.

 
Fig.1.SchematicprocedureforusingtheEpiQuikDNMT1Activity/InhibitorScreeningAssayCoreKit.

Fig.2. DNMT1inhibition:RecombinantDNMT1wasincubatedwithsubstrateandinhibitors.DNMT1activitywasmeasuredinthepresenceorabsenceofinhibitors.