TheEpiQuik™DNMT3BActivity/InhibitorScreeningAssayCoreKitisaconvenientsetoftoolsthatallowstheexperimentertoselectivelyscreenDNMT3Bactivity/inhibitorsinanELISA-likeformat.TheselectiveinhibitionofDNMT3Bmayleadtodemethylationandexpressionofsilencedtumorsuppressorgenes.Thus,theselectiveDNMT3Binhibitorscouldbeanewadditiontocancertherapeuticagents.Thekitisready-to-useandprovidesalltheessentialcomponentsneededtocarryoutasuccessfulDNMT3Bactivity/inhibitorscreeningexperimentwithouttheneedforrADIoactivityoranyspecialequipment.Thekithasthefollowingadvantages:

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

Note:This"core"kitdoesnotcomewithenzymesincluded. 

Principle&Procedure
TheEpiQuik™DNMT3BActivity/InhibitorScreeningAssayCoreKitisdesignedforscreeningDnmt3Binhibitors.Inanassaywiththiskit,theuniquecytosine-rich DNAsubstrateisstablycoatedonthestripwells.ThesewellsarespecificallytreatedtohaveahighDNAabsorptionABIlity.ADNMT3B(notincluded)enzymetransfersamethylgrouptocytosinefromAdomettomethylateDNAsubstrate.ThemethylatedDNAcanberecognizedwithananti-5-methylcytosineantibody.TheratiooramountofmethylatedDNA,whichisproportionaltoenzymeactivity,canthenbecolorimetricallyquantifiedthroughanELISA-likereaction.

 
Fig.1.SchematicprocedureforusingtheEpiQuikDNMT3BActivity/InhibitorScreeningAssayCoreKit.

Fig.2. ActivityofrecombinantDNMT3BatdifferentconcentrationswasmeasuredusingtheEpiQuik™DNMT3BActivity/InhibitorScreeningAssayCoreKit.

Fig.3. RecombinantDNMT3BwasincubatedwithsubstrateandinhibitorBD-11.DNMT3Bactivitywasmeasuredinthepresenceorabsenceofinhibitors.