INSTRUCTIONSPierceBiotechnologyPOBox117(815)968-0747www.thermoscientific.com/pierce3747N.MeridianRoadRockford,lL61105USA(815)968-7316faxNumberDescription78833NE-PERNuclearandCytoplasmicExtractionReagents,sufficientreagentsforextracting50cellpelletfractionshavingpackedcellvolumesof20µLeach(atotalof~2gcellpaste)KitContents:CytoplasmicExtractionReagentI(CERI),10mLCytoplasmicExtractionReagentII(CERII),550µLNuclearExtractionReagent(NER),5mL78835NE-PERNuclearandCytoplasmicExtractionReagents,sufficientreagentsforextracting250cellpelletfractionshavingpackedcellvolumesof20µLeach(atotalof~10gcellpaste)KitContents:CytoplasmicExtractionReagentI(CERI),50mLCytoplasmicExtractionReagentII(CERII),2.75mLNuclearExtractionReagent(NER),25mLStorage:Uponreceiptstorekitcomponentsat4°C.Productisshippedatambienttemperature.IntroductionTheThermoScientificNE-PERNuclearandCytoplasmicExtractionReagentsenablestepwiseseparationandpreparationofcytoplasmicandnuclearextractsfrommammalianculturedcellsortissue.Non-denatured,activeproteinsarepurifiedinlessthantwohours.Additionofthefirsttworeagentstoacellpelletcausescellmembranedisruptionandreleaseofcytoplasmiccontents.Afterrecoveringtheintactnucleifromthecytoplasmicextractbycentrifugation,thenucleiarelysedwithathirdreagenttoyieldthenuclearextract.Extractsobtainedwiththisproductgenerallyhavelessthan10%contaminationbetweennuclearandcytoplasmicfractions,whichissufficientpurityformostexperimentsinvolvingnuclearextracts.Nuclearextractsaregenerallypreferredtowholecelllysatesforgeneregulationstudies.Cellularcomponentspresentinwholecelllysatescanadverselyaffectnuclearproteininteractionsandstability,andnuclearproteinsaremoreconcentratedinnuclearextractsthanwholecelllysates.NuclearextractsobtainedwiththeNE-PERReagentsarecompatiblewithavarietyofdownstreamapplicationsincludingWesternblotting,theThermoScientificPierceBCAProteinAssay(ProductNo.23225),gel-shift(ProductNo.20148),reporter-geneandenzyme-activityassays.ImportantProductInformation•Useproteaseinhibi...
