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GenWay/HDAC Fluorogenic Assay Kit/GWB-4C8F93/100assays
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4000-520-616
Description:
Description:TheHDACFluorescentActivityAssaykitisacompleteassaysystemdesignedtomeasurehistonedeacetylaseactivityforscreeningandprofilingapplications.Itcomesinaconvenient96-wellformat,withallthereagentsnecessaryfor100uluorescentHDACactivitymeasurements.Inaddition,thekitincludespurifiedHDAC2andapotentHDACinhibitor,TrichostatinA,forthepositiveandnegativecontrol.TheHDACFluorescentActivityAssayisbasedontheuniquefluorogenicsubstrateandDevelopercombination.ThisassaymethodeliminatesdealingwiththerADIoactivity,extraction,andchromatographyaspectsoftraditionalassays.Usingthiskit,onlytwosimplestepsonamicrotiterplateareneededtoanalyzetheHDACactivitylevel.First,theHDACfluorometricsubstrate,comprisedofanacetylatedlysinesidechain,isincubatedwithasamplecontainingpurifiedHDACenzyme.Thedeacetylationwillsensitizethesubstrate,causingfurthertreatmentwiththeLysineDevelopertoproduceafluorophorethatcanthenbemeasuredusingafluorescencereader.COMPONENTS:FluorogenicHDACsubstrate:50ul@5mM2xHDACdeveloper:6mlHDACassaybuffer:10ulTrichostatinA:100ul@200uMOneblackplate1ugHDAC2forthepositivecontrolAPPLICATIONS:GreatforstudyingenzymekineticsandscreeningsmallmolecularinhibitorsfordrugdiscoveryandHTSapplications.STORAGE:-20°Cor-70°C.REFERENCES:1.A.Itoetal.,2001,EMBOJ.201331.2.N.A.Barlevetal.,2001,Mol.Cell81243.3.A.Itoetal.,2002,EMBOJ.216236.ASSAYPROTOCOLMaterial:Assaybuffer,BPScatalogno.50031,;Assaydeveloper,BPScatalogno.50030,;HDACSubstrate,catalognumber50037,Step1:AddallreactionmixturetoalowbindingNUNCblackplate,VWRcatalogno.62408-936,1ulofHDAC2,0.1ug/ul,39ulofHDACassaybuffer,BPScatalogno.50031,5ulof1mg/mlBSA5ulof200uMsubstrate,BPScatalogno.50037,Incubateat37°Cfor30min.Step2:StopthereactionAdd50ulofHDACassaydeveloper,2x,BPScatalogno.50030,andincubatetheplateatroomtemperaturefor15minStep3:Readsampleinamicrotiter-platereadingfluorimetercapableofexcitationatawavelengthintherangeof350-380nmanddetectionofemittedlightintherangeof440-460nm.Format:assaykit
ScientificCategory:DeacetylaseAssay
AdditionalInformation:
Name | HDACFluorogenicAssayKit |
---|---|
RelatedProductNames | histonedeacetylase,HDAC |
Purity | NA |
Format | AssayKit |
Storage | Mixedstorage |
IntendedUse | ResearchUseOnly |