Agrisera/RPB1 | RNA polymerase II subunit B1/AS11 1804/


productinformation
Background		DNA-directedRNApolymeraseIIsubunit1(EC=2.7.7.6)catalyzesthetranscriptionofDNAintoRNAusingthefourribonucleosidetriphosphatesassubstrates.ThisenzymeisthecentralcomponentofthebasalRNApolymeraseIItranscriptionmachinery.Synonymes:DNA-directedRNApolymeraseIIIlargestsubunit,DNA-directedRNApolymeraseIIsubunitRPB1.
Immunogen		KLH-conjugatedsyntheticpeptidederivedfromknownhigherplantpolymeraseIIsequences,includingArabidopsisthalianaP18616,At4g35800
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Affinitypurifiedserum
Format		Lyophilized
Quantity		50µg
Reconstitution		Forreconstitutionadd50µlofsterilewater.
Storage		storelyophilized/reconstitutedat-20°C;oncereconstitutedmakealiquotstoavoidrepeatedfreeze-thawcycles.Please,remembertospintubesbrieflypriortoopeningthemtoavoidanylossesthatmightoccurfromlyophilizedmaterialadheringtothecaporsidesofthetubes.
Testedapplications		immunoprecipitation(IP)
Relatedproducts		AS10710 \|anti-H3\|histone3,rabbitantibody collectionofantibodiestoDN/RNA/cellcycle Plantandalgalproteinextractionbuffer Secondaryantibodies
Additionalinformation		PurifiedIgGwaslyophilizedfromPBSpH7.4+0.02%sodiumazide.Thereforeonlydistilledwaterisrequiredforitsre-constitution.

applicationinformation
Recommendeddilution		1,2or5µg (IP)
Expected\|apparentMW		204kDa
Confirmedreactivity		Arabidopsisthaliana
Predictedreactivity		Amygdaluspersica,Asparagusofficinalis,Brassicarapa,Capsellarubella, Glycinemax,Hordeumvulgare,Solanumlycopersicum,Oryzaglaberrima,Panicumitalicum,Perseaamericana,Pisumsativum,Ricinuscommunis,Solanumtuberosum,Sorghumvulgare,Vitisvinifera
Notreactivein		noconfirmedexceptionsfrompredictedreactivityarecurrentlyknown
Additionalinformation
Selectedreferences		Dolataetal.(2015).NTR1isrequiredfortranscriptionelongationcheckpointsatalternativeexonsinArabidopsis.EMBOJ.2015Jan7.pii:e201489478.

applicationexample

Chromatinimmunoprecipitation(ChIP).Chromatinimmunoprecipitationwasperformedasdescribed(Bowleretal.(2004),PlantJournal)withmodifications.SonicandIPbufferwerepreparedasdescribed(Kaufmannetal.(2010)),NatureProtocols).Chromatinpelletwassonicatedat4°CwithaDiagenodeBioruptorsetathighintensityfor10min(30secon,30secoffintervals)toobtain250-500bpDNAfragments.Chromatin(20µg)wasimmunoprecipitatedwithaRNAPolIIantibody(2µg/IP)andDynabeads®ProteinG(LifeTechnologies).ReversecorsslinkingandelutionofDNAfragmentswereperformedusingChelex(Biorad)at95°C,10minasdescribed(Rowleyetal.(2013),Methods).Samplewithoutantibodywasusedtodeterminethenonspecificbackgroundpulled-downdirectlybythebeads.RelativeabundanceofregionsofinterestinimmunoprecipitatedDNAwasmeasuredbyreal-timePCR.Redlinesindicatesamplifiedregion.Valuesonthechartsareshownasthemean±SDlevel(%ofinput)ofRNAPolIIonGAPDHgene(A)andMIR168a(B)gene.Datacomesfromthreeindependentexperiments.

CourtesyofMScJakubDolata,AdamMickiewiczUniversity,Poland

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Agrisera/RPB1 | RNA polymerase II subunit B1/AS11 1804/