InputType: | HistoneExtracts |
ResearchArea: | HistoneAcetylation |
TargetApplication: | AmountQuantitation |
VesselFormat: | 96-WellPlate |
100%Guarantee: | 6months |
The EpiQuik™GlobalAcetylHistoneH3-K23QuantificationKit(Colorimetric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K23colorimetrically,usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells.Thekithasthefollowingadvantages:
- Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
- InnovativecolorimetricassaywithouttheneedforrADIoactivity,electrophoresis,orchromatography.
- SpecificallycapturesacetylH3-K23withthedetectionlimitaslowas2ng/wellanddetectionrangefrom20ng-5µg/wellofhistoneextracts.
- ThecontrolisconvenientlyincludedforthequantificationoftheamountofacetylH3-K23.
- StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
- Simple,reliable,andconsistentassayconditions.
BackgroundInformation
AcetylationofhistonesincludinghistoneH3hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransfeases(HAT)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine23(H3-K23),alongwithK9,K14andK18areprimaryacetylatedsitesofhistoneH3.AcetylationofhistoneH3-K23istightlyinvolvedinthecellcyclereguation,cellproliferationandapoptosis.AcetylationofhistoneH3-K23isalsocorrelatedwithtranscriptionactivation.AnimbalanceintheequilibriumofhistoneH3acetylationincludingK23acetylationhasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K23acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.ThusquantitativedetectionofglobalacetylhistoneH3-K23wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K23QuantificationKit(Colorimetric)providesatoolformeasuringglobalacetylationofhistoneH3-K23.
Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K23acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine23iscapturedtothestripwellscoatedwithananti-acetylH3-K23antibody.ThecapturedacetylhistoneH3-K23canthenbedetectedwithalabeleddetectionantibodyfollowedbyacolordevelopmentreagent.TheratioofacetylH3-K23isproportionaltotheintensityofabsorbance.TheabsoluteamountofacetylH3-K23canbequantifiedbycomparingtothestandardcontrol.
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.