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

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貨號: 17-10131

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Description
CatalogueNumber	17-10131
TradeName	ChIPAb+ Upstate
Description	ChIPAb+?Phospho-CREB(Ser133)-ChIPValidatedAntibodyandPrimerSet
Overview	AllChIPAb+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
BackgroundInformation	CREBisaβ-ZIPtranscriptionfactorthatactivatestargetgenesthroughcAMPresponseelements.CREBisabletomediatesignalsfromnumerousphysiologicalstimuli,resultinginregulationofabroadarrayofcellularresponses.WhileCREBisexpressedinnumeroustissues,itplaysalargeregulatoryroleinthenervoussystem.CREBisbelievedtoplayakeyroleinpromotingneuronalsurvival,precursorproliferation,neuriteoutgrowthandneuronaldifferentiationincertainneuronalpopulations.CREBpromotesoutgrowthanddifferentiationasamediatorofneurotrophinpathways.CREBisabletoselectivelyactivatenumerousdownstreamgenesthroughinteractionswithdifferentdimerizationpartners.CREBisactivatedbyphosphorylationatSer133byvarioussignalingpathwaysincludingErk,Ca2+andstresssignaling.

ProductInformation
Format	Purified
Control	IncludesnegativecontrolnormalrabbitIgGandprimersspecificforhumancFOSCRE.
Presentation	Anti-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
StorageConditions	Stablefor1yearat-20°Cfromdateofreceipt.HandlingRecommendations:Uponfirstthaw,andpriortoremovingthecap,centrifugethevialandgentlymixthesolution.Aliquotintomicrocentrifugetubesandstoreat-20°C.Avoidrepeatedfreeze/thawcycles,whichmaydamageIgGandaffectproductperformance.Note:VariABIlityinfreezertemperaturesbelow-20°Cmaycauseglycerolcontainingsolutionstobecomefrozenduringstorage.

Applications
Application	ThisChIPAb+Phospho-CREB(Ser133)-ChIPValidatedAntibody&PrimerSet(AntibodyChIP)convenientlyincludestheantibody&thespecificcontrolPCRprimers.
KeyApplications	ElectrophoreticMobilityShiftAssay Immunohistochemistry Immunoprecipitation WesternBlotting ChromatinImmunoprecipitation(ChIP)
ApplicationNotes	ChromatinImmunoprecipitation: 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
Immunogen	KLH-conjugatedsyntheticpeptidecorrespondingtotheaminoacidsincludingandencompassingphosphorylatedSer133ofCREB.
Epitope	phospho-Ser133
Concentration	PleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
Host	Rabbit
Specificity	Recognizesp43phosphorylatedCREB.Mayalsorecognizep30andp38kDaproteins.Thep30&p38proteinsmaybephosphorylatedCREMandATF-1respectively,sincethetwoproteinssharesignificanthomologytothephosphorylatedCREBimmunizingpeptide.
Isotype	IgG
SpeciesReactivity	Human Rat Mouse Hamster
AntibodyType	PolyclonalAntibody
EntrezGeneNumber	NM_134442
EntrezGeneSummary	ThisgeneencodesatranscriptionfactorthatisamemberoftheleucinezipperfamilyofDNAbindingproteins.ThisproteinbindsasahomodimertothecAMP-responsiveelement,anoctamericpalindrome.Theproteinisphosphorylatedbyseveralproteinkinases,andinducestranscriptionofgenesinresponsetohormonalstimulationofthecAMPpathway.Alternatesplicingofthisgeneresultsintwotranscriptvariantsencodingdifferentisoforms.
GeneSymbol	CREB1 CREB MGC9284
Modifications	Phosphorylation
PurificationMethod	PurifiedbyproteinA-SepharosefollowedbyimmunoadsorbantchromatographyusinganunphosphorylatedCREBaffinitygelcolumn.
UniProtNumber	P16220
UniProtSummary	FUNCTION: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
MolecularWeight	DetectsCREBonlywhenphosphorylatedonSer133(~43kDa).MayrecognizeATF-1(~38kDa)andCREM(~30kDa)asthetwoproteinssharesignificanthomologytothephosphorylatedimmunizingpeptide.

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