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Epigentek/EpiQuik循环乙酰组蛋白H3K56 ELISA试剂盒（比色法）/P-3138-48/48分析
| InputType: | PlasmaorSerum |
| ResearchArea: | HistoneMethylation |
| TargetApplication: | AmountQuantitation |
| VesselFormat: | 96-WellPlate |
| 100%Guarantee: | 6months |
The EpiQuik™CirculatingAcetylHistoneH3K56ELISAKit(Colorimetric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasurecirculatingacetylhistoneH356(H3K56ac)fromBIOLOGicalfluidsamplessuchasplasmaandserumfromhuman,mouseorrat.Thekithasthefollowingadvantages:
- Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
- Highsensitivityandspecificity.Thedetectionlimitisaslowas0.5ng/wellwithdynamicrangeof1-20ng/wellwithintheindicatedamountrangeoftheplasma/serum.OnlyrecognizesH3K56acwithnocross-reactivitywithunmodifiedH3orothermodificationsatthesamelysinesite.
- ThecontrolisconvenientlyincludedforthequantificationofH3K56ac.
- StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
- Simple,reliable,andconsistentassayconditions.
BackgroundInformation
Epigeneticactivationorinactivationofgenesplaysacriticalroleinmanyimportanthumandiseases,especiallyincancer.AmajormechanismforepigeneticgeneinactivationismethylationofCpGislandsingenomicDNAcausedbyDNAmethyltransferases.Histoneacetyltransferases(HATs)anddeacetylases(HDACs)controlorregulateDNAmethylationthroughchromatin-dependenttranscriptionrepressionoractivation.HATstransferacetylgroupsfromacetylCoAtothelysineresiduesofhistoneproteins.P300andCBParethemajorHATsthatcatalyzesacetylationofhistoneH3atlysine56(H3K56)inmammaliancells.HDAC1andSIRT1/3/6arethehistonedeacetylasesthatdeacetylateH3K56.H3K56achasbeenviewedasasignaturemarkoftranscriptionallyactivategenes,whichisplacedexclusivelyinthe5’-regiondownstreamofthepromoter.TheH3K56accanalsobechangedbyinhibitionoractivationofHATsorHDAC/SIRTs.CirculatinghistoneH3K56acinplasmaorserumhasbeenobservedanddemonstratedastheMarkerformanydifferentdiseasesorpathologicalchangessuchascancerprogression.Therefore,detectionofcirculatingH3K56acwouldprovideusefulinformationforabetterunderstandingofepigeneticregulationofgeneactivationandsilencing,histonemodification-associatedpathologicalprocesses,screeningofdisease-relatedbiomarkers,aswellasfordevelopinghistonemodification-targeteddrugs.
Principle&Procedure
TheEpiQuik™CirculatingAcetylHistoneH3K56ELISAKit(Colorimetric)isdesignedformeasuringtotalH3K56acinplasmaorserum.Inanassaywiththiskit,theHistoneH3proteinsacetylatedatK56intheplasma/serumsamplearecapturedonthestripwellscoatedwithanti-H3K56acantibody.ThecapturedH3K56acproteinscanbethenrecognizedwithdetectionantibodyfollowedbyacolordevelopmentreagent.TheratioofH3K56acisproportionaltotheintensityofabsorbance.TheabsoluteamountofH3K56accanbequantitatedbycomparingtothestandardcontrol.
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.
