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Epigentek/EpiQuik Circulating Acetyl Histone H3K27 ELISA Kit (Colorimetric)/P-3136-48/48 assays
| InputType: | PlasmaorSerum |
| ResearchArea: | HistoneMethylation |
| TargetApplication: | AmountQuantitation |
| VesselFormat: | 96-WellPlate |
| 100%Guarantee: | 6months |
The EpiQuik™CirculatingAcetylHistoneH3K27ELISAKit(Colorimetric)isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasurecirculatingacetylhistoneH327(H3K27ac)fromBIOLOGicalfluidsamplessuchasplasmaandserumfromhuman,mouseorrat.Thekithasthefollowingadvantages:
- Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
- Highsensitivityandspecificity.Thedetectionlimitisaslowas0.5ng/wellwithdynamicrangeof1-20ng/wellwithintheindicatedamountrangeoftheplasma/serum.OnlyrecognizesH3K27acwithnocross-reactivitywithunmodifiedH3orothermodificationsatthesamelysinesite.
- ThecontrolisconvenientlyincludedforthequantificationofH3K27ac.
- StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
- Simple,reliable,andconsistentassayconditions.
BackgroundInformation
Epigeneticactivationorinactivationofgenesplaysacriticalroleinmanyimportanthumandiseases,especiallyincancer.AmajormechanismforepigeneticgeneinactivationismethylationofCpGislandsingenomicDNAcausedbyDNAmethyltransferases.Histoneacetyltransferases(HATs)anddeacetylases(HDACs)controlorregulateDNAmethylationthroughchromatin-dependenttranscriptionalrepressionoractivation.HATstransferacetylgroupsfromacetylCoAtothelysineresiduesofhistoneproteins.P300isthemajorHATthatcatalyzesacetylationofhistoneH3atlysine27(H3K27)inmammaliancells.HDACsandSIRTsarethehistonedeacetylasesthatdeacetylateH3K27.H3K27achasbeenviewedasasignaturemarkoftranscriptionallyactivegenes,whichisplacedexclusivelyinthe5’-regiondownstreamofthepromoter.TheH3K27accanalsobechangedbyinhibitionoractivationofHATsorHDAC/SIRTs.CirculatinghistoneH3K27acinplasmaorserumhasbeenobservedanddemonstratedastheMarkerformanydifferentdiseasesorpathologicalchangesuchascancerprogression.Therefore,detectionofcirculatingH3K27acwouldprovideusefulinformationforabetterunderstandingofepigeneticregulationofgeneactivationandsilencing,histonemodification-associatedpathologicalprocesses,screeningofdisease-relatedbiomarkers,aswellasfordevelopinghistonemodification-targeteddrugs.
Principle&Procedure
ThiskitisdesignedformeasuringtotalH3K27acinplasmaorserum.Inanassaywiththiskit,theHistoneH3proteinsacetylatedatK27intheplasma/serumsamplearecapturedonthestripwellscoatedwithanti-H3K27acantibody.ThecapturedH3K27acproteinscanbethenrecognizedwithdetectionantibodyfollowedbyacolordevelopmentreagent.TheratioofH3K27acisproportionaltotheintensityofabsorbance.TheabsoluteamountofH3K27accanbequantitatedbycomparingtothestandardcontrol.
StartingMaterials
Inputmaterialsshouldbeplasmaorserum.Theamountofplasmaorserumforeachassaycanbe10to40µlwithanoptimalamountof30µl.
WB(10XWashBuffer)
HAB(HistoneAssayBuffer)
DS(DeveloperSolution)
SS(StopSolution)
8-WellAssayStrips(WithFrame)
ControlAssayStrips(WithFrame)#
AdhesiveCoveringFilm
DAb(DetectionAntibody,1000X)*
StandardControl(100µg/ml)
*Spinthesolutiondowntothebottompriortouse.
#ControlAssayStripsaregreentrimmedfordistinguishingfrom8-wellAssayStrips(forsamples).TheControlAssayStripsareonlyforcontroluseandshouldnotbeusedforsampleassay.
