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Millipore/17-651 | ChIPAb+ Monomethyl-Histone H4 (Lys20) - ChIP Validated Antibody and Primer Set/17-651/25 assays25 ass
  • Millipore/17-651 | ChIPAb+ Monomethyl-Histone H4 (Lys20) - ChIP Validated Antibody and Primer Set/17-651/25 assays25 ass

Millipore/17-651 | ChIPAb+ Monomethyl-Histone H4 (Lys20) - ChIP Validated Antibody and Primer Set/17-651/25 assays25 ass

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    • Description
      CatalogueNumber17-651
      BrandFamilyUpstate
      TradeName
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+Monomethyl-HistoneH4(Lys20)-ChIPValidatedAntibodyandPrimerSet
      OverviewAllChIPAb+antibodiesareindividuallyvalidatedforchromatinprecipitation,everylot,everytime.EachChIPAb+antibodysetincludescontrolprimers(testedeverylotbyqPCR)toBIOLOGicallyvalidateyourIPresultsinalocus-specificcontext.TheqPCRprotocolandprimersequencesareprovided,allowingresearcherstovalidateChIPprotocolswhenusingourantibodyintheirchromatincontext.EachsetalsoincludesanegativecontrolantibodytoensurespecificityoftheChIPreaction.
      TheChIPAb+Monomethyl-HistoneH4(Lys20)setincludestheAnti-monomethyl-HistoneH4(Lys20)antibody,anegativecontrolantibody(purifiedRabbitIgG),andqPCRprimers,whichamplifya212basepairPCRproductwithinthehumanGAPDHcodingregion.ThemonomethylhistoneH4(Lys20)andnegativecontrolantibodiesaresuppliedinascalable"perChIP"reactionsizeandcanbeusedtofunctionallyvalidatetheprecipitationofmonomethyl-histoneH4(Lys20)associatedchromatin.
      AlternateNames
      • H4K20me1
      • HistoneH4(monomethylK20)
      • H4histonefamily,memberA
      • histone1
      • H4a
      • histonecluster1
      • H4a
      ProductInformation
      FormatAffinityPurified
      Control
      • IncludesnegativecontrolrabbitIgGantibodyandcontrolprimersspecificforhumanGAPDHcodingregion.
      PresentationAnti-monomethyl-HistoneH4(Lys20)(rabbitpolyclonalIgG).Onevialcontaining100μgofaffinitypurifiedantibodyin100μL0.1MTris-Glycine(pH7.4),15mMNaCl,and0.05%NaN3.Storeat-20°C.

      NormalRabbitIgG.Onevialcontaining125μgofpurifiedrabbitIgGin125μLstoragebuffer.Storeat-20°C.

      ChIPPrimersGAPDHCodingRegion.Onevialcontaining75μLof5μMofeachprimerspecificforhumanGAPDHcodingregion.Storeat-20°C.
      FOR:GGCTCCCACCTTTCTCATCC
      REV:GGCCATCCACAGTCTTCTGG
      StorageandShippingInformation
      StorageConditionsStablefor1yearat-20°Cfromdateofreceipt.HandlingRecommendations:Uponfirstthaw,andpriortoremovingthecap,centrifugethevialandgentlymixthesolution.Aliquotintomicrocentrifugetubesandstoreat-20°C.Avoidrepeatedfreeze/thawcycles,whichmaydamageIgGandaffectproductperformance.
      Applications
      ApplicationThisChIPAb+Monomethyl-HistoneH4(Lys20)-ChIPValidatedAntibody&PrimerSetconvenientlyincludestheantibody&thespecificcontrolPCRprimers.
      KeyApplications
      • WesternBlotting
      • ChromatinImmunoprecipitation(ChIP)
      ApplicationNotesChromatinImmunoprecipitation:
      SonicatedchromatinpreparedfromHeLacells(2X106cellequivalentsperIP)wassubjectedtochromatinimmunoprecipitationusing4μgofeitheranegativecontrolantibodyorAnti-Monomethyl-HistoneH4
      (Lys20)antibodyandtheMagnaChIPAKit(Cat.#17-610).Successfulimmunoprecipitationofmonomethyl-histoneH4(Lys20)-associatedDNAfragmentswasverifiedbyqPCRusingGAPDHcodingregionChIPPrimersversusControlPrimerscorrespondingtotheGAPDHpromoter(Pleaseseefigures).DataispresentedaspercentinputofeachIPsamplerelativetoinputchromatin,withimmunoprecipitatedDNAfromnegativecontrolantibodyshownas(-)andmonomethyl-histoneH4(Lys20)shownas(+).
      PleaserefertotheEZ-MagnaChIP?A(Cat.#17-408)orEZ-ChIP?(Cat.#17-371)protocolforexperimentaldetails.

      WesternBlotAnalysis:
      0.1μg/mLofproteinswerevisualizedusingagoatanti-rabbitsecondaryantibodyconjugatedtoHRPandachemiluminescencedetectionsystem(Pleaseseefigures)

      PeptideBlockingAssay:
      40μgofhistoneH4peptidecontainingmonomethylLys20abolisheddetectionofhistoneH4byanti-monomethyl-HistoneH4(Lys20)inimmunoblotanalysisofacidextractsofHeLacells(Pleaseseefigures).
      BiologicalInformation
      ImmunogenTheimmunogenwasasyntheticpeptidecorrespondingtoaminoacids15-24ofhistoneH4,monomethylatedonLys20.
      Epitopea.a.15-24
      HostRabbit
      SpecificityRecognizeshistoneH4,Mr~11kDa.
      SpeciesReactivity
      • Human
      • GreenPlants
      • Vertebrates
      SpeciesReactivityNoteHuman.Nottestedinotherspecies.Theimmunogensequenceisidenticalinawiderangeofanimalandplantspecies,sobroadcross-reactivityisexpected.
      AntibodyTypePolyclonalAntibody
      EntrezGeneNumber
      EntrezGeneSummaryHistonesarebasicnuclearproteinsthatareresponsIBLeforthenucleosomestructureofthechromosomalfiberineukaryotes.Twomoleculesofeachofthefourcorehistones(H2A,H2B,H3,andH4)formanoctamer,aroundwhichapproximately146bpofDNAiswrappedinrepeatingunits,callednucleosomes.Thelinkerhistone,H1,interactswithlinkerDNAbetweennucleosomesandfunctionsinthecompactionofchromatinintohigherorderstructures.ThisgeneisintronlessandencodesamemberofthehistoneH4family.TranscriptsfromthisgenelackpolyAtailsbutinsteadcontainapalindromicterminationelement.Thisgeneisfoundinthelargehistonegeneclusteronchromosome6.
      GeneSymbol
      • H4/A
      • H4/B
      • H4/C
      • H4/D
      • H4/E
      • H4/G
      • H4/H
      • H4/I
      • H4/J
      • H4/J
      • H4/M
      • H4/N
      • H4/O
      • H4/a
      • H4F2
      • H4FA
      • H4FB
      • H4FC
      • H4FD
      • H4FE
      • H4FG
      • H4FH
      • H4FI
      • H4FJ
      • H4FK
      • H4FM
      • H4FN
      • H4FO
      • HIST2H4
      Modifications
      • Methylation
      UniProtNumber
      UniProtSummaryFUNCTION:Corecomponentofnucleosome.NucleosomeswrapandcompactDNAintochromatin,limitingDNAaccessibilitytothecellularmachinerieswhichrequireDNAasatemplate.Histonestherebyplayacentralroleintranscriptionregulation,DNArepair,DNAreplicationandchromosomalstABIlity.DNAaccessibilityisregulatedviaacomplexsetofpost-translationalmodificationsofhistones,alsocalledhistonecode,andnucleosomeremodeling.

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

      SUBCELLULARLOCATION:Nucleus.

      PTM:AcetylationatLys-6,Lys-9,Lys-13andLys-17occursincodingregionsofthegenomebutnotinheterochromatin.

      CitrullinationatArg-4byPADI4impairsmethylation.

      MonomethylationatArg-4byPRMT1favorsacetylationatLys-9andLys-13.DemethylationisperformedbyJMJD6.

      Monomethylated,dimethylatedortrimethylatedatLys-21.MonomethylationisperformedbySET8.TrimethylationisperformedbySUV420H1andSUV420H2andinducesgenesilencing.

      UbiquitinatedbytheCUL4-DDB-RBX1complexinresponsetoultravioletirradiation.ThismayweakentheinteractionbetweenhistonesandDNAandfacilitateDNAaccessibilitytorepairproteins.

      Sumoylated,whichisassociatedwithtranscriptionalrepression.

      SEQUENCESIMILARITY:BelongstothehistoneH4family.

      SEQUENCECAUTION:ThesequenceAAI28106.1differsfromthatshown.Reason:Frameshiftatposition3.

      MolecularWeight~11kDa
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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