The EpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Fluorometric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K9fluorometrically, usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells. Thekithasthefollowingadvantages:

• Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
• InnovativefluorometricassaywithouttheneedforrADIoactivity,electrophoresis,orchromatography.
• SpecificallycapturesacetylH3-K9withthedetectionlimitaslowas0.4ng/wellanddetectionrangefrom5ng-2µg/wellofhistoneextracts.
• ThecontrolisconvenientlyincludedforthequantificationoftheamountofacetylH3-K9.
• StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
• Simple,reliable,andconsistentassayconditions.

BackgroundInformation
Acetylationofhistones,includinghistoneH3,hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransferases(HAT)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine9(H3-K9)isaprimaryacetylatedsiteofhistoneH3.AcetylationofhistoneH3-K9appearstohaveadominantroleinhistonedepositionandchromatinassemblyinsomeorganisms.AcetylationofhistoneH3-K9istightlyinvolvedinthecellcycleregulation,cellproliferation,andapoptosis.AcetylationofhistoneH3-K9isalsoanactiveMarker,andthebalancebetweenhistoneH3-K9acetylationandmethylationisimportantfortheestablishmentofspecificchromatinstructures.AnimbalanceintheequilibriumofhistoneH3acetylation,includingK9acetylation,hasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K9acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.Thus,quantitativedetectionofglobalacetylhistoneH3-K9wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Fluorometric)providesatoolformeasuringglobalacetylationofhistoneH3-K9.

Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K9acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine9iscapturedtothestripwellscoatedwithananti-acetylH3-K9antibody.ThecapturedacetylhistoneH3-K9canthenbedetectedwithalabeleddetectionantibodyfollowedbyafluorescentdevelopmentreagent.TheratioofacetylH3-K9isproportionaltotheintensityoffluorescence.TheabsoluteamountofacetylH3-K9canbequantifiedbycomparingtothestandardcontrol.

StartingMaterials
Inputmaterialshouldbepurifiedhistoneextracts.Ingeneral,theinputamountshouldbefrom50ngto200ngperwellofhistoneextracts.

F1(10Xwashbuffer)
F2(antibodybuffer)
F3(detectionantibody,1mg/ml)*
F4(fluorodeveloper)*
F5(fluoroenhancer)*
F6(fluorodilution)
Standardcontrol(100µg/ml)*
8wellsamplestrips(withframe)
8wellstandardcontrolstrips
Userguide

*Formaximumrecoveryoftheproducts,centrifugetheoriginalvialpriortoopeningthecap.