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Millipore/17-10131 | ChIPAb+™ Phospho-CREB (Ser133) - ChIP Validated Antibody and Primer Set/17-10131/25 assays25 assays
  • Millipore/17-10131 | ChIPAb+™ Phospho-CREB (Ser133) - ChIP Validated Antibody and Primer Set/17-10131/25 assays25 assays

Millipore/17-10131 | ChIPAb+™ Phospho-CREB (Ser133) - ChIP Validated Antibody and Primer Set/17-10131/25 assays25 assays

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货号: 17-10131
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    • Description
      CatalogueNumber17-10131
      TradeName
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™Phospho-CREB(Ser133)-ChIPValidatedAntibodyandPrimerSet
      OverviewAllChIPAb+antibodiesareindividuallyvalidatedforchromatinprecipitation,everylot,everytime.EachChIPAb+antibodysetincludescontrolprimers(testedeverylotbyqPCR)toBIOLOGicallyvalidateyourIPresultsinalocus-specificcontext.TheqPCRprotocolandprimersequencesareprovided,allowingresearcherstovalidateChIPprotocolswhenusingourantibodyintheirchromatincontext.EachsetalsoincludesanegativecontrolantibodytoensurespecificityoftheChIPreaction.
      TheChIPAb+Phospho-CREB(Ser133)setincludesthePhospho-CREB(Ser133)antibody,anegativecontrolnormalrabbitIgG,andqPCRprimerswhichamplifya64bpregionofhumancFosCRE.ThePhospho-CREB(Ser133)andnegativecontrolsaresuppliedinascalable"perChIP"reactionsizeandcanbeusedtofunctionallyvalidatetheprecipitationofPhospho-CREB(Ser133)-associatedchromatin.
      AlternateNames
      • activetranscriptionfactorCREB
      • cAMPresponsiveelementbindingprotein1
      • cAMP-responseelement-bindingprotein-1
      • transactivatorprotein
      BackgroundInformationCREBisaβ-ZIPtranscriptionfactorthatactivatestargetgenesthroughcAMPresponseelements.CREBisabletomediatesignalsfromnumerousphysiologicalstimuli,resultinginregulationofabroadarrayofcellularresponses.WhileCREBisexpressedinnumeroustissues,itplaysalargeregulatoryroleinthenervoussystem.CREBisbelievedtoplayakeyroleinpromotingneuronalsurvival,precursorproliferation,neuriteoutgrowthandneuronaldifferentiationincertainneuronalpopulations.CREBpromotesoutgrowthanddifferentiationasamediatorofneurotrophinpathways.CREBisabletoselectivelyactivatenumerousdownstreamgenesthroughinteractionswithdifferentdimerizationpartners.CREBisactivatedbyphosphorylationatSer133byvarioussignalingpathwaysincludingErk,Ca2+andstresssignaling.
      ProductInformation
      FormatPurified
      Control
      • IncludesnegativecontrolnormalrabbitIgGandprimersspecificforhumancFOSCRE.
      PresentationAnti-phospho-CREB(Ser133)(rabbitpolyclonal),PartNo.CS204400.Onevialcontaining100µgofaffinitypurifiedrabbitserumin0.1MTris-Glycine(pH7.4)0.15MNaCl,and0.05%sodiumazide,beforetheadditionof30%glycerol.Storeat-20°C.
      NormalRabbitIgG,PartNo.PP64B.Onevialcontaining125µgRabbitIgGin125µLstoragebuffercontaining0.05%sodiumazide.
      Storeat-20°C.
      ChIPPrimerscFosCRE,PartNo.CS203203.Onevialcontaining75μLof5μMofeachprimerspecificforthecFosCRE.Storeat-20°C.
      FOR:GGCCCACGAGACCTCTGAGACA
      REV:GCCTTGGCGCGTGTCCTAATCT
      StorageandShippingInformation
      StorageConditionsStablefor1yearat-20°Cfromdateofreceipt.HandlingRecommendations:Uponfirstthaw,andpriortoremovingthecap,centrifugethevialandgentlymixthesolution.Aliquotintomicrocentrifugetubesandstoreat-20°C.Avoidrepeatedfreeze/thawcycles,whichmaydamageIgGandaffectproductperformance.Note:VariABIlityinfreezertemperaturesbelow-20°Cmaycauseglycerolcontainingsolutionstobecomefrozenduringstorage.
      Applications
      ApplicationThisChIPAb+Phospho-CREB(Ser133)-ChIPValidatedAntibody&PrimerSet(AntibodyChIP)convenientlyincludestheantibody&thespecificcontrolPCRprimers.
      KeyApplications
      • ElectrophoreticMobilityShiftAssay
      • Immunohistochemistry
      • Immunoprecipitation
      • WesternBlotting
      • ChromatinImmunoprecipitation(ChIP)
      ApplicationNotesChromatinImmunoprecipitation:
      Representativelotdata.
      Sonicatedchromatinpreparedfrom293Tcells(5X10E6cellequivalentsperIP)weresubjectedtochromatinimmunoprecipitationusingeither4µgofNormalRabbitIgG,(PartNo.P64B),or4µgofAnti-Phospho-CREB(Ser133)antibody(PartNo.CS204400)andtheMagnaChIP™AKit(Cat.#17-610).
      SuccessfulimmunoprecipitationofPhospho-CREB(Ser133)antibodyassociatedDNAfragmentswasverifiedbyqPCRusingChIPPrimerscFosCRE(PartNo.CS203203)asapositivelocus,andcFosdownstream4Kbasanegativelocus(Figure2).DataispresentedaspercentinputofeachIPsamplerelativetoinputchromatinforeachampliconandChIPsampleasindicated.
      PleaserefertotheEZ-MagnaChIP™A(Cat.#17-408)orEZ-ChIP™(Cat.#17-371)protocolforexperimentaldetails.
      WesternBlotAnalysis:
      Representativelotdata.
      Untreated(lane1)andForskolin-treated(Lane2)NIH/3T3lysateswereresolvedbySDS-PAGE,transferredtoPVDF,andprobedwithanti-phospho-CREB(Ser133)(1:1,000dilution).
      Proteinswerevisualizedusingadonkeyanti-rabbitsecondaryantibodyconjugatedtoHRPandachemiluminescencedetectionsystem.
      ArrowindicatesphosphorylatedCREB(Figure3).
      Immunoprecipitation:
      10µLfromarepresentativelotofthisantibodywasshowntoimmunoprecipitatephosphorylatedCREBfrom500µgofcellstreatedwith50mMforskolin.
      Immunohistochemistry:A1:1000dilutionofarepresentativelotdetectedphosphorylatedCREBinformalin-fixedparaffin-embeddednormalandischemicratbrainsections.
      ElectrophoreticMobilityShift:
      Arepresentativelotofthisantibodyhasbeenshowntogelshiftbyanindependentlaboratory.
      BiologicalInformation
      ImmunogenKLH-conjugatedsyntheticpeptidecorrespondingtotheaminoacidsincludingandencompassingphosphorylatedSer133ofCREB.
      Epitopephospho-Ser133
      ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
      HostRabbit
      SpecificityRecognizesp43phosphorylatedCREB.Mayalsorecognizep30andp38kDaproteins.Thep30&p38proteinsmaybephosphorylatedCREMandATF-1respectively,sincethetwoproteinssharesignificanthomologytothephosphorylatedCREBimmunizingpeptide.
      IsotypeIgG
      SpeciesReactivity
      • Human
      • Rat
      • Mouse
      • Hamster
      AntibodyTypePolyclonalAntibody
      EntrezGeneNumber
      EntrezGeneSummaryThisgeneencodesatranscriptionfactorthatisamemberoftheleucinezipperfamilyofDNAbindingproteins.ThisproteinbindsasahomodimertothecAMP-responsiveelement,anoctamericpalindrome.Theproteinisphosphorylatedbyseveralproteinkinases,andinducestranscriptionofgenesinresponsetohormonalstimulationofthecAMPpathway.Alternatesplicingofthisgeneresultsintwotranscriptvariantsencodingdifferentisoforms.
      GeneSymbol
      • CREB1
      • CREB
      • MGC9284
      Modifications
      • Phosphorylation
      PurificationMethodPurifiedbyproteinA-SepharosefollowedbyimmunoadsorbantchromatographyusinganunphosphorylatedCREBaffinitygelcolumn.
      UniProtNumber
      UniProtSummaryFUNCTION:ThisproteinbindsthecAMPresponseelement(CRE),asequencepresentinmanyviralandcellularpromoters.CREBstimulatestranscriptiononbindingtotheCRE.TransciptionactivationisenhancedbytheTORCcoactivatorswhichactindependentlyofSer-133phosphorylation.ImplicatedinsynchronizationofcircADIanrhythmicity.
      SUBUNIT:InteractswithPPRC1.BindsDNAasadimer.Thisdimerisstabilizedbymagnesiumions.Interacts,throughthebZIPdomain,withthecoactivatorsTORC1/CRTC1,TORC2/CRTC2andTORC3/CRTC3.WhenphosphorylatedonSer-133,bindsCREBBPBysimilarity.
      SUBCELLULAR:Nucleus.
      PMT:Stimulatedbyphosphorylation.PhosphorylationofbothSer-133andSer-142intheSCNregulatestheactivityofCREBandparticipatesincircadianrhythmgeneration.PhosphorylationofSer-133allowsCREBBPbindingBysimilarity.PhosphorylateduponDNAdamage,probablybyATMorATR.
      SIMILARITY:BelongstothebZIPfamily.
      Contains1bZIPdomain.
      Contains1KID(kinase-inducIBLe)domain
      MolecularWeightDetectsCREBonlywhenphosphorylatedonSer133(~43kDa).MayrecognizeATF-1(~38kDa)andCREM(~30kDa)asthetwoproteinssharesignificanthomologytothephosphorylatedimmunizingpeptide.
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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