
- MultiSpecies
- USE-MeasureallHDMs;LSD1andJumonjitype
- SAMPLE-Lysates,PurifiedEnzymeSystems
- SAMPLES/KIT-89inDuplicate
- CALIBRATED-MeasureFormaldehydeProducedbyDemethylation
- STABILITY-Stable4˚CLiquidReagents
TheDetectX®UniversalHistoneDemethylase(HDM)FluorescentActivitykitallowsallknownHDMstobemeasuredinlow,mediumandhighthroughputfashion.RunthedemethylasereactionandaftercompletionsimplyaddthesuppliedFormaldehydeDetectionReagenttoeachwellandreadthefluorescentsignalgenerated.Excitationat450nm/Emissionat510nmusinganadjustablegainplatefluorimeter.
Formaldehydeisacommonbyproductformedinoxidativedemethylation.Examplesofformaldehyde-producingenzymesincludehistonedemethylases(HDMs)thatmodifymethylatedhistones.Lysine-specificHDMswerefirstdiscoveredin2004andarecurrentlyamongthemostactivelystudiedformaldehyde-producingenzymes.HDMscatalyzethesite-specificdemethylationofmethyl-lysineresiduesinhistonestodynamicallyregulatechromatinstructure,geneexpression,andpotentiallyothergenomicfunctions.Atpresent,therearetwoknownclassesofHDMs:theflavinadeninenucleotide(FAD)-dependentLysineSpecificDemethylase1(LSD1)familyandtheFe(II)-dependentJumonjiC(JmjC)family.AlthoughtheLSD1andJmjCHDMsemploydifferentcofactorsandcatalyticmechanisms,bothproduceformaldehydeasabyproductofthedemethylationreaction.DespitetheirBIOLOGicalimportance,HDMshaveprovendifficulttoquantitativelyassayowingtotheirrelativelylowturnovernumbers,hinderingourunderstandingoftheirkineticproperties,substratespecificities,andreactionmechanisms.
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