InputType: | HistoneExtracts |
ResearchArea: | HistoneAcetylation |
TargetApplication: | AmountQuantitation |
VesselFormat: | 96-WellPlate |
100%Guarantee: | 6months |
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.