The EpiQuik™GlobalAcetylHistoneH3-K14QuantificationKit(Colorimetric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K14colorimetrically,usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells.Thekithasthefollowingadvantages:

• Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
• InnovativecolorimetricassaywithouttheneedforrADIoactivity,electrophoresis,orchromatography.
• SpecificallycapturesacetylH3-K14withthedetectionlimitaslowas2ng/wellanddetectionrangefrom20ng-5µg/wellofhistoneextracts.
• ThecontrolisconvenientlyincludedforthequantificationoftheamountofacetylH3-K14.
• StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
• Simple,reliable,andconsistentassayconditions.

BackgroundInformation
Acetylationofhistones,includinghistoneH3,hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransferases(HATs)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine14(H3-K14),alongwithK9andK18areprimaryacetylatedsitesofhistoneH3.AcetylationofhistoneH3-K14istightlyinvolvedinthecellcycleregulation,cellproliferation,andapoptosis.AcetylationofhistoneH3-K14isalsocorrelatedwithtranscriptionactivation.AnimbalanceintheequilibriumofhistoneH3acetylation,includingK14acetylation,hasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K14acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.Thus,quantitativedetectionofglobalacetylhistoneH3-K14wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivation,andfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K14QuantificationKit(Colorimetric)providesatoolformeasuringglobalacetylationofhistoneH3-K14.

Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K14acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine14iscapturedtothestripwellscoatedwithananti-acetylH3-K14antibody.ThecapturedacetylhistoneH3-K14canthenbedetectedwithalabeleddetectionantibodyfollowedbyacolordevelopmentreagent.TheratioofacetylH3-K14isproportionaltotheintensityofabsorbance.TheabsoluteamountofacetylH3-K14canbequantifiedbycomparingtothestandardcontrol.

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.