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