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Millipore/17-10054 | ChIPAb+ Histone H2B - ChIP Validated Antibody and Primer Set/17-10054/25 assays25 assays per set. R
  • Millipore/17-10054 | ChIPAb+ Histone H2B - ChIP Validated Antibody and Primer Set/17-10054/25 assays25 assays per set. R

Millipore/17-10054 | ChIPAb+ Histone H2B - ChIP Validated Antibody and Primer Set/17-10054/25 assays25 assays per set. R

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    • Description
      CatalogueNumber17-10054
      BrandFamilyUpstate
      TradeName
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+HistoneH2B-ChIPValidatedAntibodyandPrimerSet
      OverviewAllChIPAb+antibodiesareindividuallyvalidatedforchromatinprecipitation,everylot,everytime.EachChIPAb+antibodysetincludescontrolprimers(testedeverylotbyqPCR)toBIOLOGicallyvalidateyourIPresultsinalocus-specificcontext.TheqPCRprotocolandprimersequencesareprovided,allowingresearcherstovalidateChIPprotocolswhenusingourantibodyintheirchromatincontext.EachsetalsoincludesanegativecontrolantibodytoensurespecificityoftheChIPreaction.
      TheChIPAb+HistoneH2BsetincludestheHistoneH2Bantibody,anegativecontrolrabbitIgG,andqPCRprimerswhichamplifya110bpregionofhumanβ-globinpromoter.TheHistoneH2Bandnegativecontrolsaresuppliedinascalable"perChIP"reactionsizeandcanbeusedtofunctionallyvalidatetheprecipitationofHistoneH2B-associatedchromatin.
      AlternateNames
      • H2B
      • HistoneH2B
      • H2Bhistonefamily,memberQ
      • histone2,H2be
      • histonecluster2,H2be
      BackgroundInformationHistoneH2Bisoneofthe5mainhistoneproteinsinvolvedinthestructureofchromatinineukaryoticcells.FeaturingamainglobulardomainandalongNterminaltailH2Aisinvolvedwiththestructureofthenucleosomesofthe"beadsonastring"structure.
      ProductInformation
      FormatPurified
      Control
      • IncludesnegativecontrolnormalrabbitIgGantibodyandprimersspecificforhumanβ-globin.
      PresentationAnti-HistoneH2B(rabbitpolyclonal).Onevialcontaining25μLofproteinApurifiedrabbitpolyclonalinbuffercontaining0.02Mphosphatebuffer,pH7.6,0.25MNaCl,and0.1%sodiumazide,and30%glycerol.Storeat-20°C.

      NormalRabbitIgG.Onevialcontaining125μgofpurifiedrabbitIgGin125μLstoragebuffer.Storeat-20°C.

      ChIPPrimers,β-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:VariABIllityinfreezertemperaturesbelow-20°Cmaycauseglycerolcontainingsolutionstobecomefrozenduringstorage.
      Applications
      ApplicationTheChIPAb+HistoneH2BsetincludestheHistoneH2Bantibody,anegativecontrolrabbitIgG&qPCRprimerswhichamplifya110bpregionofhumanβ-globinpromoter.
      KeyApplications
      • WesternBlotting
      • ChromatinImmunoprecipitation(ChIP)
      ApplicationNotesChromatinImmunoprecipitation:
      SonicatedchromatinpreparedfromHeLacells(1X106cellequivalentsperIP)weresubjectedtochromatinimmunoprecipitationusingeither1μLofNormalRabbitIgGor1μLofAnti-HistoneH2BandtheMagnaChIP?AKit(Cat.#17-610).
      SuccessfulimmunoprecipitationofHistoneH2BassociatedDNAfragmentswasverifiedbyqPCRusingChIPPrimers,humanβ-globinforapositive,andhumanGAPDHprimersasanegativeassay(Pleaseseefigures).
      DataispresentedaspercentinputofeachIPsamplerelativetoinputchromatinforeachampliconandChIPsampleasindicated.
      PleaserefertotheEZ-MagnaChIP?A(Cat.#17-408)orEZ-ChIP?(Cat.#17-371)protocolforexperimentaldetails.

      WesternBlotAnalysis:
      Representativelotdata.
      RecombinantXenopusHistoneH2B,chickencorehistonesandacidextractfromeithersodiumbutyrateorcolcemidtreatedHeLacellswereresolvedbyelectrophoresis,transferredtonitrocelluloseandprobedwithanti-HistoneH2B(1:5,000dilution).
      Proteinswerevisualizedusingagoatanti-rabbitsecondaryantibodyconjugatedtoHRPandachemiluminescencedetectionsystem(Pleaseseefigures).
      BiologicalInformation
      ImmunogenKLH-conjugated,syntheticpeptidecorrespondingtoaminoacids118-126(CG-AVTKYTSSK)ofhumanHistoneH2B,withanN-terminalCGaddedforconjugationpurposes.
      HostRabbit
      SpecificityRecognizesandisspecificforHistoneH2B,MW~15kDa.Anadditionalbandat~23kDaisdetectedinsomesamples,whichlikelycorrespondstoubiquityl-HistoneH2B.
      SpeciesReactivity
      • Chicken
      • Xenopus
      • Human
      • Yeast(S.cerevisiae)
      SpeciesReactivityNoteHuman,chicken,yeastandXenopus.Othersnottested.Broadspeciescross-reactivityexpectedbasedonsequencehomology.
      AntibodyTypePolyclonalAntibody
      EntrezGeneNumber
      EntrezGeneSummaryHistonesarebasicnuclearproteinsthatareresponsIBLeforthenucleosomestructureofthechromosomalfiberineukaryotes.Nucleosomesconsistofapproximately146bpofDNAwrappedaroundahistoneoctamercomposedofpairsofeachofthefourcorehistones(H2A,H2B,H3,andH4).Thechromatinfiberisfurthercompactedthroughtheinteractionofalinkerhistone,H1,withtheDNAbetweenthenucleosomestoformhigherorderchromatinstructures.ThisgeneisintronlessandencodesamemberofthehistoneH2Bfamily.Transcriptsfromthisgenecontainapalindromicterminationelement.
      GeneSymbol
      • GL105
      • H2B
      • H2B-GL105
      • H2B.1
      • H2B.q
      • H2B/q
      • H2BFQ
      • MGC119802
      • MGC119804
      • MGC129733
      • MGC129734
      • OTTHUMP00000013920
      PurificationMethodProteinAPurfied
      UniProtNumber
      UniProtSummaryFUNCTION:SwissProt:Q8N257#Corecomponentofnucleosome.NucleosomeswrapandcompactDNAintochromatin,limitingDNAaccessibilitytothecellularmachinerieswhichrequireDNAasatemplate.Histonestherebyplayacentralroleintranscriptionregulation,DNArepair,DNAreplicationandchromosomalstability.DNAaccessibilityisregulatedviaacomplexsetofpost-translationalmodificationsofhistones,alsocalledhistonecode,andnucleosomeremodeling.
      SIZE:126aminoacids;13908Da
      SUBUNIT:ThenucleosomeisahistoneoctamercontainingtwomoleculeseachofH2A,H2B,H3andH4assembledinoneH3-H4heterotetramerandtwoH2A-H2Bheterodimers.Theoctamerwrapsapproximately147bpofDNA.
      SUBCELLULARLOCATION:Nucleus.
      PTM:MonoubiquitinationofLys-121bytheRNF20/40complexgivesaspecifictagforepigenetictranscriptionalactivationandisalsoprerequisiteforhistoneH3"Lys-4"and"Lys-79"methylation.ItalsofunctionscooperativelywiththeFACTdimertostimulateelongationbyRNApolymeraseII.&PhosphorylatedonSer-15bySTK4/MST1duringapoptosis;whichfacilitatesapoptoticchromatincondensation.AlsophosphorylatedonSer-15inresponsetoDNAdoublestrandbreaks(DSBs),andincorrelationwithsomatichypermutationandimmunoglobulinclass-switchrecombination.
      SIMILARITY:SwissProt:Q8N257##BelongstothehistoneH2Bfamily.
      MolecularWeight~15kDa
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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