The EpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Colorimetric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K9colorimetrically,usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells. Thekithasthefollowingadvantages:

• Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
• InnovativecolorimetricassaywithouttheneedforrADIoactivity,electrophoresis,orchromatography.
• SpecificallycapturesacetylH3-K9withthedetectionlimitaslowas2ng/wellanddetectionrangefrom20ng-5µg/wellofhistoneextracts.
• ThecontrolisconvenientlyincludedforthequantificationoftheamountofacetylH3-K9.
• StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
• Simple,reliable,andconsistentassayconditions.

BackgroundInformation
Acetylationofhistones,includinghistoneH3,hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransferases(HATs)andhistonedeacetylases(HDACs)playacriticalroleinthecontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine9(H3-K9)isaprimaryacetylatedsiteofhistoneH3.AcetylationofhistoneH3-K9appearstohaveadominantroleinhistonedepositionandchromatinassemblyinsomeorganisms.AcetylationofhistoneH3-K9istightlyinvolvedincellcycleregulation,cellproliferation,andapoptosis;acetylationofhistoneH3-K9isalsoanactiveMarker.ThebalancebetweenhistoneH3-K9acetylationandmethylationisimportantfortheestablishmentofspecificchromatinstructures.AnimbalanceintheequilibriumofhistoneH3acetylation,includingK9acetylation,hasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K9acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.Thus,quantitativedetectionofglobalacetylhistoneH3-K9wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Colorimetric)providesatoolformeasuringglobalacetylationofhistoneH3-K9.

Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K9acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine9iscapturedtothestripwellscoatedwithananti-acetylH3-K9antibody.ThecapturedacetylhistoneH3-K9canthenbedetectedwithalabeleddetectionantibodyfollowedbyacolordevelopmentreagent.TheratioofacetylH3-K9isproportionaltotheintensityofabsorbance.TheabsoluteamountofacetylH3-K9canbequantifiedbycomparingtothestandardcontrol.

StartingMaterials
Inputmaterialshouldbepurifiedhistoneextracts.Ingeneral,theinputamountshouldbefrom50ngto200ngperwellofhistoneextracts. 

C1(10Xwashbuffer)
C2(antibodybuffer)
C3(detectionantibody,1mg/ml)*
C4(colordeveloper)
C5(stopsolution)
Standardcontrol(100µg/ml)*
8wellsamplestrips(withframe)
8wellstandardcontrolstrips
Userguide

*Formaximumrecoveryoftheproducts,centrifugetheoriginalvialpriortoopeningthecap.