GenWay/LSD1 Fluorescent Assay kit/GWBF41821/100reactions

Description:

Description:TheLSD1AssayKitisdesignedtodetectLSD1activityusingfluorimetry.TheassayisbasedontheanalysisofthehydrogeneperoxideproducedduringthedemethylationreactionmediatedbyLSD1.Humanlysine-specificdemethylase,LSD1,isachromatin-modifyingenzymethatspecificallyremovesmethylgroupsfrommono-anddimethylatedLysofhistoneH3.LSD1isapotentialtargetfordrugdevelopment.TheLSD1AssayKitcomesinaconvenient96-wellformat,withpurifiedLSD1,AmplexRedreagent,peroxidase,andLSD1assaybufferfor100enzymereactions.ThekeytotheLSD1AssayKitisthemethylatedpeptidesubstrate.Usingthiskit,onlytwosimplestepsonamicrotiterplatearerequiredforLSD1reactions.First,asamplecontainingLSD1enzymeisincubatedinareactionmixturewiththemethylatedpeptidesubstrate.Second,peroxidaseandAmplexRedreagentareaddedandfluorescence,?exc=530+/-13nm,?em=590+/-18nm,ismeasuredusingaplatereader.Components:LSD120µg,700µMmethylatedpeptidesubstrate1ml,10mMAmplexRedreagent100µl,2xLSD1assaybuffer20ul,10U/mlperoxidase200µl,96-wellmoduleAPPLICATIONS:GreatforstudyingenzymekineticsandscreeningsmallmolecularinhibitorsfordrugdiscoveryandHTSapplications.STABILITY:1yearwhenstoredaslisted.REFERENCES:1.FornerisF,BindaC,Dall,AglioA,FraaijeMW,BattaglioliE,andMatteviA.JBiolChem2006;281,46,:35289-95.2.ZhouM,DiwuZ,Panchuk-VoloshinaN,andHauglandRP.AnalBiochem1997;253,2,:162-8.AssayProtocol:Materials:2xassaybuffer,methylatedpeptidesubstrate,BPScatalognumber50101,LSD1,BPScatalognumber50100,AmplexRedreagent,peroxidaseStep1:1,Addtheentirereactionmixturetoa96-wellmodule,VWRcatalogno.62409-300,:40µlLSD1,5ng/μl;dilutewith1xLSD1assaybuffer,10µlofmethylatedpeptidesubstrate,700µM,30µlof2xLSD1assaybuffer20µlH2O2,Incubatefor15-30minatroomtemperature.Step2:1,Immediatelybeforeuse:add1µlof10mMAmplexRedstockand2µlof10U/mlperoxidaseto97µlof1xLSD1buffer,mix.2,Add100µlofmixturetoeachwell.Mixwell.Incubateatroomtemperaturefor2-4minutes.3,Readfluorescenceat?exc=530nmand?em=590nm.

Format:assaykit

ScientificCategory:Demethylase/Assay

AdditionalInformation:

Name	LSD-1FluorescentAssaykit
RelatedProductNames	LSD-1FluorescentAssaykit
Format	AssayKit
Storage	Mixedstorage
IntendedUse	ResearchUseOnly