InputType: | HistoneExtracts |
ResearchArea: | HistoneAcetylation |
TargetApplication: | AmountQuantitation |
VesselFormat: | 96-WellPlate |
100%Guarantee: | 6months |
The EpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Fluorometric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K9fluorometrically, usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells. Thekithasthefollowingadvantages:
BackgroundInformation
Acetylationofhistones,includinghistoneH3,hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransferases(HAT)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine9(H3-K9)isaprimaryacetylatedsiteofhistoneH3.AcetylationofhistoneH3-K9appearstohaveadominantroleinhistonedepositionandchromatinassemblyinsomeorganisms.AcetylationofhistoneH3-K9istightlyinvolvedinthecellcycleregulation,cellproliferation,andapoptosis.AcetylationofhistoneH3-K9isalsoanactiveMarker,andthebalancebetweenhistoneH3-K9acetylationandmethylationisimportantfortheestablishmentofspecificchromatinstructures.AnimbalanceintheequilibriumofhistoneH3acetylation,includingK9acetylation,hasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K9acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.Thus,quantitativedetectionofglobalacetylhistoneH3-K9wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Fluorometric)providesatoolformeasuringglobalacetylationofhistoneH3-K9.
Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K9acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine9iscapturedtothestripwellscoatedwithananti-acetylH3-K9antibody.ThecapturedacetylhistoneH3-K9canthenbedetectedwithalabeleddetectionantibodyfollowedbyafluorescentdevelopmentreagent.TheratioofacetylH3-K9isproportionaltotheintensityoffluorescence.TheabsoluteamountofacetylH3-K9canbequantifiedbycomparingtothestandardcontrol.
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.
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