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

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貨號: 17-651

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Description
CatalogueNumber	17-651
BrandFamily	Upstate
TradeName	ChIPAb+ Upstate
Description	ChIPAb+Monomethyl-HistoneH4(Lys20)-ChIPValidatedAntibodyandPrimerSet
Overview	AllChIPAb+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
Format	AffinityPurified
Control	IncludesnegativecontrolrabbitIgGantibodyandcontrolprimersspecificforhumanGAPDHcodingregion.
Presentation	Anti-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
StorageConditions	Stablefor1yearat-20°Cfromdateofreceipt.HandlingRecommendations:Uponfirstthaw,andpriortoremovingthecap,centrifugethevialandgentlymixthesolution.Aliquotintomicrocentrifugetubesandstoreat-20°C.Avoidrepeatedfreeze/thawcycles,whichmaydamageIgGandaffectproductperformance.

Applications
Application	ThisChIPAb+Monomethyl-HistoneH4(Lys20)-ChIPValidatedAntibody&PrimerSetconvenientlyincludestheantibody&thespecificcontrolPCRprimers.
KeyApplications	WesternBlotting ChromatinImmunoprecipitation(ChIP)
ApplicationNotes	ChromatinImmunoprecipitation: 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
Immunogen	Theimmunogenwasasyntheticpeptidecorrespondingtoaminoacids15-24ofhistoneH4,monomethylatedonLys20.
Epitope	a.a.15-24
Host	Rabbit
Specificity	RecognizeshistoneH4,Mr~11kDa.
SpeciesReactivity	Human GreenPlants Vertebrates
SpeciesReactivityNote	Human.Nottestedinotherspecies.Theimmunogensequenceisidenticalinawiderangeofanimalandplantspecies,sobroadcross-reactivityisexpected.
AntibodyType	PolyclonalAntibody
EntrezGeneNumber	NM_003538.3
EntrezGeneSummary	HistonesarebasicnuclearproteinsthatareresponsIBLeforthenucleosomestructureofthechromosomalfiberineukaryotes.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	P62805
UniProtSummary	FUNCTION: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

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