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

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貨號: 17-10250

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Description
CatalogueNumber	17-10250
TradeName	ChIPAb+ Upstate
Description	ChIPAb+?Dimethyl-HistoneH4(Arg3)Symmetric-ChIPValidatedAntibodyandPrimerSet
Overview	AllChIPAb+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
BackgroundInformation	HistonesarehighlyconservedproteinsthatserveasthestructuralscaffoldfortheorganizationofnuclearDNAintochromatin.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
Format	AffinityPurified
Control	IncludesnormalrabbitIgGandprimersspecificforhumanβ-globin.
Presentation	Anti-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
StorageConditions	Stablefor1yearat-20°Cfromdateofreceipt.HandlingRecommendations:Uponfirstthaw,andpriortoremovingthecap,centrifugethevialandgentlymixthesolution.Aliquotintomicrocentrifugetubesandstoreat-20°C.Avoidrepeatedfreeze/thawcycles,whichmaydamageIgGandaffectproductperformance. Note:VariABIlityinfreezertemperaturesbelow-20°Cmaycauseglycerolcontainingsolutionstobecomefrozenduringstorage.

Applications
Application	ThisChIPAb+Dimethyl-HistoneH4(Arg3)Symmetric-ChIPValidatedAntibody&PrimerSetconvenientlyincludestheantibody&thespecificcontrolPCRprimers.
KeyApplications	DotBlot WesternBlotting ChromatinImmunoprecipitation(ChIP)
ApplicationNotes	ChromatinImmunoprecipitation: 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
Immunogen	KLH-conjugatedlinearpeptidecorrespondingtohumanHistoneH4whensymmetricallydimethylatedatArg3.
Epitope	DimethylatedArg3
Concentration	PleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
Host	Rabbit
Specificity	ThisantibodyrecognizesHistoneH4symmetricallydimethylatedatArg3.
SpeciesReactivity	Human Mouse Rat
SpeciesReactivityNote	Demonstratedtoreactwithhuman,mouse,andrat.Broadspeciescross-reactivityexpectedbasedonsequencesimilarity.
AntibodyType	PolyclonalAntibody
EntrezGeneNumber	NP_001029249
EntrezGeneSummary	HistonesarebasicnuclearproteinsthatareresponsIBLeforthenucleosomestructureofthechromosomalfiberineukaryotes.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
PurificationMethod	AffinityPurfied
UniProtNumber	P62805
UniProtSummary	FUNCTION: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.

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