ThePARPtrap™AssayKitisdesignedtomeasurePARP1/DNAcomplexformationinahighthroughputscreeningassayusingfluorescencepolarization(FP).ThePARPtrap™AssayKitcomesinaconvenient96-wellformat,withpurifiedPARP1enzyme,fluorescentlabelednickedDNA,andPARPtrap™assaybufferfor100enzymereactions.ThekeytothePARPtrap™AssayKitisthefluorescentlabelednickedDNA.WithoutthePARreaction,PARP1bindstothefluorescentlabelednickedDNA,resultingintheemissionofhighlypolarizedlight.However,afterautoribosylationofPARP1,thenickedDNAisdissociatedfromPARP1androtatesfreely,emittinglesspolarizedlight(Fig.1).
Figure1.PARPtrap™AssayKitschematic
COMPONENTS:
1.Murai,J.,etal.MolecularCancerTherapeutics2014.13:433-443.
2.Murai,J.,et.al.CancerResearch2012.72:5588-5599.
MATERIALSORINSTRUMENTSREQUIREDBUTNOTSUPPLIED:
Fluorescentmicroplatereadercapableofmeasuringfluorescencepolarization
Adjustablemicropipettorandsteriletips
Rotatingorrockerplatform