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Epigentek/EpiQuik Circulating Acetyl Histone H3K18 ELISA Kit (Colorimetric)/P-3134-48/48 assays
| InputType: | PlasmaorSerum |
| ResearchArea: | HistoneMethylation |
| TargetApplication: | AmountQuantitation |
| VesselFormat: | 96-WellPlate |
| 100%Guarantee: | 6months |
The EpiQuik™CirculatingAcetylHistoneH3K18ELISAKit(Colorimetric)isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasurecirculatingacetylhistoneH3K18(H3K18ac)fromBIOLOGicalfluidsamplessuchasplasmaandserumfromhuman,mouseorrat.Thekithasthefollowingadvantages:
- Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
- Highsensitivityandspecificity.Thedetectionlimitisaslowas0.5ng/wellwithdynamicrangeof1-20ng/wellwithintheindicatedamountrangeoftheplasma/serum.OnlyrecognizesH3K18acwithnocross-reactivitywithunmodifiedH3orothermodificationsatthesamelysinesite.
- ThecontrolisconvenientlyincludedforthequantificationofH3K18ac.
- StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
- Simple,reliable,andconsistentassayconditions.
BackgroundInformation
Epigeneticactivationorinactivationofgenesplaysacriticalroleinmanyimportanthumandiseases,especiallyincancer.AmajormechanismforepigeneticgeneinactivationismethylationofCpGislandsingenomicDNAcausedbyDNAmethyltransferases.Histoneacetyltransferases(HATs)anddeacetylases(HDACs)controlorregulateDNAmethylationthroughchromatin-dependenttranscriptionalrepressionoractivation.HATstransferacetylgroupsfromacetylCoAtothelysineresiduesofhistoneproteins.P300isthemajorHATthatcatalyzesacetylationofhistoneH3atlysine18(H3K18)inmammaliancells.HDACsandSIRT1/7arethemajorhistonedeacetylasesthatdeacetylateH3K18.H3K18achasbeenviewedasasignaturemarkoftranscriptionallyactivegenes,whichisplacedexclusivelyinthe5’-regiondownstreamofthepromoter.TheH3K18accanalsobechangedbyinhibitionoractivationofHATsorHDAC/SIRTs.CirculatinghistoneH3K18acinplasmaorserumhasbeenobservedanddemonstratedastheMarkerformanydifferentdiseasesorpathologicalchangessuchascancerprogression.Therefore,detectionofcirculatingH3K18acwouldprovideusefulinformationforabetterunderstandingofepigeneticregulationofgeneactivationandsilencing,histonemodification-associatedpathologicalprocesses,screeningofdisease-relatedbiomarkers,aswellasfordevelopinghistonemodification-targeteddrugs.
Principle&Procedure
ThiskitisdesignedformeasuringtotalH3K18acinplasmaorserum.Inanassaywiththiskit,theHistoneH3proteinsacetylatedatK18intheplasma/serumsamplearecapturedonthestripwellscoatedwithanti-H3K18acantibody.ThecapturedH3K18acproteinscanbethenrecognizedwithdetectionantibodyfollowedbyacolordevelopmentreagent.TheratioofH3K18acisproportionaltotheintensityofabsorbance.TheabsoluteamountofH3K18accanbequantitatedbycomparingtothestandardcontrol.
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.
