InputType: | HistoneExtracts |
ResearchArea: | HistoneCitrullination |
TargetApplication: | AmountQuantitation |
VesselFormat: | 96-WellPlate |
100%Guarantee: | 6months |
TheEpiQuik™HistoneH3CitrullinationELISAKit (Colorimetric) isacompletesetofoptimizedbuffersandreagentsforspecificallymeasuringglobalhistoneH3citrullinationfromabroadrangeofspeciessuchasmammals,fungi,andbacteria,inavarietyofformsincludingculturedcellsandfreshtissues.Histoneextractscanbepreparedbyusingyourownsuccessfulmethod.Foryourconvenienceandthebestresults,Epigentekoffersahistoneextractionkitoptimizedforusewiththiskit.Histoneextractscanbeusedimmediatelyorstoredat–80°Cforfutureuse.Thiskithasthefollowingadvantages:
BackgroundInformation
Argininehistonecitrullinationisoneofthemanyimportantepigeneticmarks,andisessentialfortheregulationofepigeneticchromatinremodelingandimmunecell’sextracellulartrapprocesses.CitrullinatedarginineofhistoneH3(Arg2,8,17)regulatesgeneexpressionandismainlycatalyzedbypeptidylargininedeiminase4(PAD4).FurThermore,citrullinationofhistones,inparticularhistoneH3,wasrevealedasaconvergencepointfordiverseinflammatorysignalsthattriggertheneutrophilresponsetoinfections.ItwasreportedthatcitrullinatedhistoneH3couldbeapotentialserumbioMarkerfortheearlydiagnosisofsepticshock.MoreimmunologicaldiseasessuchasmultiplesclerosisandrheumatoidarthritisseemtobealsoassociatedwithchangeofhistoneH3citrullination.TheglobalhistoneH3citrullinationcanbechangedbyinhibitionoractivationofPADs.Therefore,quantitativedetectionofglobalhistoneH3citrullinationwouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandsilencing,aswellasfordevelopingPAD-targeteddrugs.
Principle&Procedure
ThiskitisdesignedformeasuringglobalH3cit.Inanassaywiththiskit,thehistoneproteinsarestablyspottedonthestripwells.ThecitrullineH3canberecognizedwithahigh-affinityantibodyanddetectedwithadetectionantibody,followedbyacolordevelopmentreagent.TheratioofH3citisproportionaltotheintensityofabsorbance.TheabsoluteamountofH3citcanbequantitatedbycomparingtothestandardcontrol.
StartingMaterials
Inputmaterialsshouldbehistoneextracts.Theamountofhistoneextractsforeachassaycanbe50ngto200ngwithanoptimalamountof100ng.
WB(10XWashBuffer)
BB(BlockingBuffer)
DS(DeveloperSolution)
SS(StopSolution)
HCB(HistoneCoatingBuffer)
8-WellAssayStrips(WithFrame)
ControlStrips(WithFrame)#
AdhesiveCoveringFilm
CAb(CaptureAntibody,1000X)*
DAb(DetectionAntibody,1000X)*
H3citControl(50µg/ml)
*Spinthesolutiondowntothebottompriortouse.
#ControlStripsaregreentrimmedfordistinguishingfrom8-wellAssayStrips(forsamples).
TheControlStripsareonlyforcontroluseandshouldnotbeusedforsampleassay.
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