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Millipore/17-10250 | ChIPAb+™ Dimethyl-Histone H4 (Arg3) Symmetric - ChIP Validated Antibody and Primer Set/17-10250/25
  • Millipore/17-10250 | ChIPAb+™ Dimethyl-Histone H4 (Arg3) Symmetric - ChIP Validated Antibody and Primer Set/17-10250/25

Millipore/17-10250 | ChIPAb+™ Dimethyl-Histone H4 (Arg3) Symmetric - ChIP Validated Antibody and Primer Set/17-10250/25

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货号: 17-10250
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    • Description
      CatalogueNumber17-10250
      TradeName
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™Dimethyl-HistoneH4(Arg3)Symmetric-ChIPValidatedAntibodyandPrimerSet
      OverviewAllChIPAb+antibodiesareindividuallyvalidatedforchromatinprecipitation,everylot,everytime.EachChIPAb+antibodysetincludescontrolprimers(testedeverylotbyqPCR)toBIOLOGicallyvalidateyourIPresultsinalocus-specificcontext.TheqPCRprotocolandprimersequencesareprovided,allowingresearcherstovalidateChIPprotocolswhenusingourantibodyintheirchromatincontext.EachsetalsoincludesanegativecontrolantibodytoensurespecificityoftheChIPreaction.
      TheChIPAb+Dimethyl-HistoneH4(Arg3)SymmetricsetincludestheDimethyl-HistoneH4(Arg3)Symmetricantibody,aNormalRabbitIgG,andcontrolprimerswhichamplifya110bpregionofChIPPrimers,humanβ-globin.TheDimethyl-HistoneH4(Arg3)Symmetricandnegativecontrolsaresuppliedinascalable"perChIP"reactionsizeandcanbeusedtofunctionallyvalidatetheprecipitationofDimethyl-HistoneH4(Arg3)Symmetric-associatedchromatin.
      AlternateNames
      • HistoneH4
      BackgroundInformationHistonesarehighlyconservedproteinsthatserveasthestructuralscaffoldfortheorganizationofnuclearDNAintochromatin.Thefourcorehistones,H2A,H2B,H3,andH4,assembleintoanoctamer(2moleculesofeach).Histonesaremodifiedpost-translationallybytheactionsofenzymesinboththenucleusandcytoplasm.Thesemodifications,whichincludeacetylation,phosphorylation,methylation,andubiquitin-ation,occurpredominantlyontheN-terminalandC-terminaltailsthatextendbeyondthenucleosomecoreparticle.Argininemodificationsofhistonesproteinsaremediatedbytwoclassesofproteinargininemethyltransferases(PRMTs),classIandII.ClassIPRMTsplacetwomethylgroupsontheonenitrogenatomofthearginineguanidinogroup,resultinginanasymmetricmodification.IncontrastclassIIPRMTsplacetwomethylgroupsontwonitrogenatomsoftheguanidinogroupproducingasymmetricdimethylation.Althoughthereareexceptions,asymmetricmodificationsaretypicallyassociatedwithtranscriptionactivationwhilesymmetricmodificationsaremostcommonlyassociatedwithtranscriptionalrepression.
      ProductInformation
      FormatAffinityPurified
      Control
      • IncludesnormalrabbitIgGandprimersspecificforhumanβ-globin.
      PresentationAnti-Dimethyl-HistoneH4(Arg3)Symmetric(rabbitpolyclonal).Onevialcontaining17.5µgofpurifiedrabbitpolyclonalinbuffercontaining0.1MTris-Glycine(pH7.4),150mMNaClwith0.05%sodiumazidebeforetheadditionofglycerolto30%.Storeat-20°C.
      Concentration:0.7µg/µL
      NormalRabbitIgG.Onevialcontaining125µgofRabbitIgGin125µLofstoragebuffercontaining0.05%sodiumazide.Storeat-20°C.
      ChIPPrimers,humanβ-globin.Onevialcontaining75μLof5μMofeachprimerspecificforthehumanβ-globinpromoter.Storeat-20°C.
      FOR:AGGACAGGTACGGCTGTCATC
      REV:TTTATGCCCAGCCCTGGCTC
      StorageandShippingInformation
      StorageConditionsStablefor1yearat-20°Cfromdateofreceipt.HandlingRecommendations:Uponfirstthaw,andpriortoremovingthecap,centrifugethevialandgentlymixthesolution.Aliquotintomicrocentrifugetubesandstoreat-20°C.Avoidrepeatedfreeze/thawcycles,whichmaydamageIgGandaffectproductperformance.
      Note:VariABIlityinfreezertemperaturesbelow-20°Cmaycauseglycerolcontainingsolutionstobecomefrozenduringstorage.
      Applications
      ApplicationThisChIPAb+Dimethyl-HistoneH4(Arg3)Symmetric-ChIPValidatedAntibody&PrimerSetconvenientlyincludestheantibody&thespecificcontrolPCRprimers.
      KeyApplications
      • DotBlot
      • WesternBlotting
      • ChromatinImmunoprecipitation(ChIP)
      ApplicationNotesChromatinImmunoprecipitation:
      Representativelotdata.
      SonicatedchromatinpreparedfromHeLacells(1X10E6cellequivalentsperIP)weresubjectedtochromatinimmunoprecipitationusing1µgofeitherNormalrabbitIgGor0.7µgofAnti-Dimethyl-HistoneH4(Arg3)SymmetricandtheMagnaChIP™AKit(Cat.#17-610).SuccessfulimmunoprecipitationofDimethyl-HistoneH4(Arg3)SymmetricassociatedDNAfragmentswasverifiedbyqPCRusingChIPPrimers,humanβ-globinasapositivelocus,andGAPDHpromoterprimers(22-004)asanegativelocus.(Figure2).DataispresentedaspercentinputofeachIPsamplerelativetoinputchromatinforeachampliconandChIPsampleasindicated.
      PleaserefertotheEZ-MagnaChIP™A(Cat.#17-408)orEZ-ChIP™(Cat.#17-371)protocolforexperimentaldetails.
      WesternBlotAnalysis:
      Representativelotdata.
      HeLaacidextract(Lane1)andrecombinantHistoneH4(Lane2)wereprobedwithAnti-Dimethyl-HistoneH4(Arg3)Symmetrical(0.1μg/mL).ProteinswerevisualizedusingaDonkeyAnti-RabbitIgGsecondaryantibodyconjugatedtoHRPandachemiluminescencedetectionsystem.
      ArrowindicatesHistoneH4(~13kDa).(Figure3).
      Note:Anuncharacterizedbandat~58kDahasbeenobservedinsomelysates.
      DotBlotAnalysis:
      Representativelotdata.
      Histonepeptideswithvariousmodifications(seetablebelow)wereprobedwithAnti-Dimethyl-HistoneH4(Arg3)Symmetrical(1:2,000dilution).ProteinswerevisualizedusingaDonkeyAnti-RabbitIgGsecondaryantibodyconjugatedtoHRPandachemiluminescencedetectionsystem.(Figure4).Seetablebelow.
      ImmunocytochemistryAnalysis:
      Representativelotdata.
      ConfocalfluorescentanalysisofA431andHeLacellsusingAnti-Dimethyl-HistoneH4(Arg3)Symmetrical(Red).ActinfilamentshavebeenlabeledwithAlexaFluor®488dyePhalloidin(Green).NucleusisstainedwithDAPI(Blue).Thisantibodypositivelystainsthenucleus.(Figure5).
      BiologicalInformation
      ImmunogenKLH-conjugatedlinearpeptidecorrespondingtohumanHistoneH4whensymmetricallydimethylatedatArg3.
      EpitopeDimethylatedArg3
      ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
      HostRabbit
      SpecificityThisantibodyrecognizesHistoneH4symmetricallydimethylatedatArg3.
      SpeciesReactivity
      • Human
      • Mouse
      • Rat
      SpeciesReactivityNoteDemonstratedtoreactwithhuman,mouse,andrat.Broadspeciescross-reactivityexpectedbasedonsequencesimilarity.
      AntibodyTypePolyclonalAntibody
      EntrezGeneNumber
      EntrezGeneSummaryHistonesarebasicnuclearproteinsthatareresponsIBLeforthenucleosomestructureofthechromosomalfiberineukaryotes.Thisstructureconsistsofapproximately146bpofDNAwrappedaroundanucleosome,anoctamercomposedofpairsofeachofthefourcorehistones(H2A,H2B,H3,andH4).Thechromatinfiberisfurthercompactedthroughtheinteractionofalinkerhistone,H1,withtheDNAbetweenthenucleosomestoformhigherorderchromatinstructures.ThisgeneisintronlessandencodesamemberofthehistoneH4family.TranscriptsfromthisgenelackpolyAtails;instead,theycontainapalindromicterminationelement.Thisgeneisfoundinahistoneclusteronchromosome1.Thisgeneisoneoffourhistonegenesintheclusterthatareduplicated;thisrecordrepresentsthetelomericcopy.[providedbyRefSeq].
      GeneSymbol
      • HIST1H4A
      • H4/J
      • HIST1H4F
      • HIST1H4L
      • H4FN
      • H4FA
      • H4FH
      • HIST1H4H
      • H4F2
      • H4FM
      • H4/A
      • H4FK
      • H4FG
      • HIST2H4
      • H4FB
      • H4/K
      • HIST1H4I
      • H4/N
      • HIST1H4B
      • H4FE
      • H4/M
      • H4/a
      • HIST1H4E
      • HIST4H4
      • HIST2H4A
      • H4FC
      • H4FI
      • H4/H
      • HIST1H4C
      • H4/G
      • HIST1H4K
      • H4FJ
      • H4FD
      • H4/I
      • H4/B
      • H4/D
      • H4/C
      • HIST1H4J
      • HIST1H4D
      • H4/E
      Modifications
      • Methylation
      PurificationMethodAffinityPurfied
      UniProtNumber
      UniProtSummaryFUNCTION:Corecomponentofnucleosome.NucleosomeswrapandcompactDNAintochromatin,limitingDNAaccessibilitytothecellularmachinerieswhichrequireDNAasatemplate.Histonestherebyplayacentralroleintranscriptionregulation,DNArepair,DNAreplicationandchromosomalstability.DNAaccessibilityisregulatedviaacomplexsetofpost-translationalmodificationsofhistones,alsocalledhistonecode,andnucleosomeremodeling.

      SUBUNITSTRUCTURE:ThenucleosomeisahistoneoctamercontainingtwomoleculeseachofH2A,H2B,H3andH4assembledinoneH3-H4heterotetramerandtwoH2A-H2Bheterodimers.Theoctamerwrapsapproximately147bpofDNA.

      SUBCELLULARLOCATION:Nucleus.Chromosome.

      PTM:AcetylationatLys-6(H4K5ac),Lys-9(H4K8ac),Lys-13(H4K12ac)andLys-17(H4K16ac)occursincodingregionsofthegenomebutnotinheterochromatin.

      CitrullinationatArg-4(H4R3ci)byPADI4impairsmethylation.

      MonomethylationatArg-4(H4R3me1)byPRMT1favorsacetylationatLys-9(H4K8ac)andLys-13(H4K12ac).DemethylationisperformedbyJMJD6.

      SymmetricdimethylationonArg-4(H4R3me2a)bythePRDM1/PRMT5complexmayplayacrucialroleinthegerm-celllineage.

      Monomethylated,dimethylatedortrimethylatedatLys-21(H4K20me1,H4K20me2,H4K20me3).MonomethylationisperformedbySET8.TrimethylationisperformedbySUV420H1andSUV420H2andinducesgenesilencing.

      UbiquitinatedbytheCUL4-DDB-RBX1complexinresponsetoultravioletirradiation.ThismayweakentheinteractionbetweenhistonesandDNAandfacilitateDNAaccessibilitytorepairproteins.MonoubiquitinatedatLys-92ofhistoneH4(H4K91ub1)inresponsetoDNAdamage.TheexactroleofH4K91ub1inDNAdamageresponseisstillunclearbutitmayfunctionasalicensingsignalforadditionalhistoneH4post-translationalmodificationssuchasH4Lys-21methylation(H4K20me).

      Sumoylated,whichisassociatedwithtranscriptionalrepression.

      SEQUENCESIMILARITIES:BelongstothehistoneH4family.

      SEQUENCECAUTION:ThesequenceAAI28106.1differsfromthatshown.Reason:Frameshiftatposition3.
      MolecularWeight~13kDaobserved.Crossreactingbandat~58kDahasbeenobservedinsomelysates.
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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