InputType: | HistoneExtracts |
ResearchArea: | HistoneAcetylation |
TargetApplication: | AmountQuantitation |
VesselFormat: | 96-WellPlate |
100%Guarantee: | 6months |
The EpiQuik™GlobalAcetylHistoneH3-K14QuantificationKit(Fluorometric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K14fluorometrically,usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells.Thekithasthefollowingadvantages:
BackgroundInformation
AcetylationofhistonesincludinghistoneH3hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransfeases(HAT)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine14(H3-K14),alongwithK9andK18areprimaryacetylatedsitesofhistoneH3.AcetylationofhistoneH3-K14istightlyinvolvedinthecellcyclereguation,cellproliferationandapoptosis.AcetylationofhistoneH3-K14isalsocorrelatedwithtranscriptionactivation.AnimbalanceintheequilibriumofhistoneH3acetylationincludingK14acetylationhasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K14acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.ThusquantitativedetectionofglobalacetylhistoneH3-K14wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K14QuantificationKit(Fluorometric)providesatoolformeasuringglobalacetylationofhistoneH3-K14.
Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K14acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine14iscapturedtothestripwellscoatedwithananti-acetylH3-K14antibody.ThecapturedacetylhistoneH3-K14canthenbedetectedwithalabeleddetectionantibodyfollowedbyafluorescentdevelopmentreagent.TheratioofacetylH3-K14isproportionaltotheintensityoffluorescence.TheabsoluteamountofacetylH3-K14canbequantifiedbycomparingtothestandardcontrol.
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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