TheFluorogenicHDAC5AssayKitisacompleteassaysystemdesignedtomeasurehistonedeacetylase5(HDAC5)activityforscreeningandprofilingapplications.Itcomesinaconvenient96-wellformat,withallthereagentsnecessaryfor100fluorescentHDAC5activitymeasurements.Inaddition,thekitincludespurifiedHDAC5enzymeandapotentHDACinhibitor,TrichostatinA,foruseasapositiveandnegativecontrol.TheFluorogenicHDAC5AssayKitisbasedonauniquefluorogenicsubstrateanddevelopercombination.Thisassaymethodeliminatesdealingwithther
ADIoactivity,extraction,andchromatographyaspectsoftraditionalassays.Usingthiskit,onlytwosimplestepsonamicrotiterplateareneededtoanalyzetheHDAC5activitylevel.First,theHDACfluorometricsubstrate,containinganacetylatedlysinesidechain,isincubatedwithpurifiedHDAC5.ThedeacetylationsensitizesthesubstratesosubsequenttreatmentwiththeLysineDeveloperproducesafluorophorethatcanthenbemeasuredusingafluorescencereader.