The EpiQuik™GlobalAcetylHistoneH3-K18QuantificationKit(Fluorometric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K18 fluorometrically,usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells.Thekithasthefollowingadvantages:

• Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
• InnovativefluorometricassaywithouttheneedforrADIoactivity,electrophoresis,orchromatography.
• SpecificallycapturesacetylH3-K18withthedetectionlimitaslowas0.4ng/wellanddetectionrangefrom5ng-2µg/wellofhistoneextracts.
• ThecontrolisconvenientlyincludedforthequantificationoftheamountofacetylH3-K18.
• StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
• Simple,reliable,andconsistentassayconditions.

BackgroundInformation
Acetylationofhistones,includinghistoneH3,hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransferases(HAT)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine18(H3-K18)-alongwithK9andK14-areprimaryacetylatedsitesofhistoneH3.AcetylationofhistoneH3-K18istightlyinvolvedinthecellcycleregulation,cellproliferation,andapoptosis.AcetylationofhistoneH3-K18isalsocorrelatedwithtranscriptionactivation.AnimbalanceintheequilibriumofhistoneH3acetylation,includingK18acetylation,hasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K18acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.Thus,quantitativedetectionofglobalacetylhistoneH3-K18wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K18QuantificationKit(Fluorometric)providesatoolformeasuringglobalacetylationofhistoneH3-K18.

Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K18acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine18iscapturedtothestripwellscoatedwithananti-acetylH3-K18antibody.ThecapturedacetylhistoneH3-K18canthenbedetectedwithalabeleddetectionantibodyfollowedbyafluorescentdevelopmentreagent.TheratioofacetylH3-K18isproportionaltotheintensityoffluorescence.TheabsoluteamountofacetylH3-K18canbequantifiedbycomparingtothestandardcontrol.

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.