TheEpiQuik™TotalHistoneExtractionHTKitprovidesasimpleandhighthroughputmicroplatemethodforextractinghistoneproteinsusedforavarietyofapplications,whichincludehistonemodificationssuchasacetylation,methylation,andsumoylation.ThiskitisalsospecificallydesignedtomeettherequirementsofhistoneextractsusedinEpiQuik™histonequantificationassays.Thekitcanbeusedtodirectlyextracthistonesfrommammaliancellsculturedina96-wellplate.TheEpiQuik™TotalHistoneExtractionHTKithasthefastestprocedureavailableonthemarket,allowingcompletionwithin60minutes. - Pre-optimizedandsimple1hourprotocol.
- Convenientlyincludesallessentialreagentstocarryoutahistoneextraction.
- StandardizedprocedureforreproducIBLeresults.
- Extractsahighyieldoftotalcorehistonesfromaslittleas 104 cells.
- Post-translationalmodificationsarekeptintact.
- Doesnotaffecthistonemodificationstatusorlevels.
BackgroundInformation
HistonesarethechiefproteincomponentsofchromatininBIOLOGy.TheyactasspoolsaroundwhichDNAwinds,andalsoplayaroleingeneregulation.ThecorehistonesincludeH2A,H2B,H3,andH4.Histonesundergopost-translationalmodifications,whichaltertheirinteractionwithDNAandnuclearproteins.TheH3andH4histoneshavelongtailsprotrudingfromthenucleosome,whichcanbecovalentlymodifiedatseveralplaces.Modificationsofthetailincludemethylation,acetylation,phosphorylation,ubiquitination,sumoylation,citrullination,andADP-ribosylation(H2Acanalsobemodified).Combinationsofmodificationsarethoughttoconstituteacode,theso-called"histonecode."Histonemodificationsactindiversebiologicalprocessessuchasgeneregulation,DNArepairandchromosomecondensation(mitosis). Principle&Procedure
TheEpiQuik™TotalHistoneExtractionHTKitsimplyappliesourproprietaryhistoneisolationbufferstocells.AftertreatmentwithPre-lysis,Lysis,andBalancebuffers,thetotalhistonesareeasilyextractedforimmediateuseorstorageatproperconditions. StartingMaterials
Theminimalamountofstartingmaterialscanbeaslowas 104 cells.Forthebestresults,thecellnumbershouldbegreaterthan3x104 cells.Atotalof2x96standardextractions(use3x104 cells)canbeperformedwiththiskit. | 
 Fig.1.SchematicprocedureforusingtheEpiQuik™TotalHistoneExtractionHTKit.
Fig.2. HistoneextractswerepreparedfromMCF-7cells(30,000cells/well)usingtheEpiQuikTotalHistoneExtractionHTKitandacetylhistoneH3wasquantifiedusingthe EpiQuikGlobalPan-MethylHistoneH3K9QuantificationKit(Colorimetric)(Cat.#P-3036).
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