Epigentek/EpiQuik Global Acetyl Histone H3K18 Quantification Kit (Colorimetric)/P-4014-48/48 assays

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

• Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
• InnovativecolorimetricassaywithouttheneedforrADIoactivity,electrophoresis,orchromatography.
• SpecificallycapturesacetylH3-K18withthedetectionlimitaslowas2ng/wellanddetectionrangefrom20ng-5µg/wellofhistoneextracts.
• ThecontrolisconvenientlyincludedforthequantificationoftheamountofacetylH3-K18.
• StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
• Simple,reliable,andconsistentassayconditions.

BackgroundInformation
Acetylationofhistones,includinghistoneH3,hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransferases(HATs)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine18(H3-K18),alongwithK9andK14,areprimaryacetylatedsitesofhistoneH3.AcetylationofhistoneH3-K18istightlyinvolvedinthecellcyclereguation,cellproliferationandapoptosis.AcetylationofhistoneH3-K18isalsocorrelatedwithtranscriptionactivation.AnimbalanceintheequilibriumofhistoneH3acetylation,includingK18acetylation,hasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K18acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.Thus,quantitativedetectionofglobalacetylhistoneH3-K18wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivation,andfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K18QuantificationKit(Colorimetric)providesatoolformeasuringglobalacetylationofhistoneH3-K18.

Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K18acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine18iscapturedtothestripwellscoatedwithananti-acetylH3-K18antibody.ThecapturedacetylhistoneH3-K18canthenbedetectedwithalabeleddetectionantibody,followedbyacolordevelopmentreagent.TheratioofacetylH3-K18isproportionaltotheintensityofabsorbance.TheabsoluteamountofacetylH3-K18canbequantifiedbycomparingtothestandardcontrol.

StartingMaterials
Inputmaterialshouldbepurifiedhistoneextracts.Ingeneral,theinputamountshouldbefrom50ngto200ngperwellofhistoneextracts.

C1(10Xwashbuffer)
C2(antibodybuffer)
C3(detectionantibody,1mg/ml)*
C4(colordeveloper)
C5(stopsolution)
Standardcontrol(100µg/ml)*
8-Wellsamplestrips(withframe)
8-Wellstandardcontrolstrips
UserGuide

*Formaximumrecoveryoftheproducts,centrifugetheoriginalvialpriortoopeningthecap.