TheEpiQuik™TotalHistoneExtractionKit isacompletesetofoptimizedbuffersandreagentsforextractingtotalcorehistoneproteins(H2A,H2B,H3,andH4)frommammaliancellsortissues inasimple60minuteprocedure. Thepost-translationalmodifications(PTM)inthehistoneextractsarekeptintactandthuscanbeusedwithEpigentek"s histonemodification assaykits orinavarietyofdownstreamapplicationsforhistone methylation,acetylation,phosphorylation, sumoylation,ubiquitination, citrullination,andADP-ribosylation  studies.

• Pre-optimizedandsimple1hourprotocol.
• Convenientlyincludesallessentialreagentstocarryoutahistoneextraction.
• StandardizedprocedureforreproducIBLeresults.
• Extractsahighyieldoftotalcorehistonesfromaslittleas 1 mgoftissues.
• Post-translationalmodificationsarekeptintact.
• Doesnotaffecthistonemodificationstatusorlevels.

BackgroundInformation
HistonesarethechiefproteincomponentsofchromatininBIOLOGy.TheyactasspoolsaroundwhichDNAwinds,andalsoplayaroleingeneregulation.ThecorehistonesincludeH2A,H2B,H3,andH4.Histonesundergopost-translationalmodifications,whichaltertheirinteractionwithDNAandnuclearproteins.TheH3andH4histoneshavelongtailsprotrudingfromthenucleosome,whichcanbecovalentlymodifiedatseveralplaces(i.e.,methylation,acetylation,phosphorylation,etc.).Histonemodificationsactindiversebiologicalprocessessuchasgeneregulation,DNArepairandchromosomecondensation(mitosis).Combinationsofmodificationsarethoughttoconstitutea "histonecode"andanunderstandingofthiscodemayhelptoanswerquestionsabouthumandevelopmentanddisease.

Principle&Procedure
TheEpiQuik™TotalHistoneExtractionKitsimplyappliesourproprietaryhistoneisolationbufferstocellortissuesamples.Aftertreatmentwithpre-lysis,lysis,andbalancebuffers,thetotalcore histonesareeasilyextractedforimmediateuseorstorageunderproperconditions.

StartingMaterials
Theminimalamountofstartingmaterialscanbeaslowas10⁵ cellsor1mgtissue.Forthebestresults,thecellnumbershouldbegreaterthan10⁶ cellsorthetissueamountshouldbegreaterthan10mg.Atotalof100standardextractions(using10⁷ cellsor100mgtissuesperextraction)canbeperformedwiththiskit.Yieldofthetotalhistoneproteinsisapproximately0.4mgper10⁷ cellsor100mgoftissue.Theyieldmayvarydependingoncellortissuetype.

10XPre-LysisBuffer
LysisBuffer
BalanceBuffer
DTTSolution
UserGuide