ProductDescription
Size:100μl
AbType:Polyclonal
Reactivity:Eukaryotes
Immunogen:Di-methylated H3(Lys27)-KLH
Host:Rabbit
ProductDescriptionThisproductisproducedbyimmunizingrabbitswithasyntheticdi-methylpeptidecorrespondingtoresiduessurroundingLys27ofhumanhistoneH3.AntibodiesarepurifiedbyproteinA-conjugatedagarosefollowedbydi-methylatedhistoneH3(Lys27)peptideaffinitychromatography.
SpecificityThisantibodydetectshistoneH3onlywhenitisdi-methylatedatLys27,butnotthemono-,di-methylatedandthenon-methylatedH3peptideatLys27;mono-methylatedhistoneH3atLys23.
RecommendedApplicationsChIP,IP.Nottestedinotherapplications*IP=ImmunoprecipitationChIP=Chromatinprecipitation
RecommendedantibodydilutionIP:1::25ChIP:1:25(4μlofantibodyfor1x105HeLa)
ImportantNotesUseatanassaydependentconcentration.Optimaldilutions/concentrationsshouldbedeterminedbytheenduser.
A.Dotblottinganalysisonindicatedamountofdi-methylatedH3peptideatLys27(lane1),mono-methylatedH3peptideatLys27(lane2),tri-methylatedH3peptideatLys27(lane3),mono-methylatedH3peptideatLys23(lane4)andnon-methylatedH3peptideatLys27(lane5)usingdi-methyl-histoneH3(Lys27)rabbitpAb.
B.Sonicatedchromatinpreparedfrom1x105HeLacellswassubjectedtochromatinimmunoprecipitation(ChIP)usingeither4μlDi-methyl-HistoneH3(Lys27)rabbitpAbor1μgofnormalrabbitIgGasanegativecontrol.RealtimequantitativePCRwasperformedonimmunoprecipitatedDNAusingprimersspecificforthehumanRPL30Exon3andMyoD1Exon1.DataarepresentedasenrichmentofeachIPedsamplerelativetototalamountofinputchromatinateachamplicon.
ScientificDescriptionTheε-aminolysineacetylationofproteinsisanimportantrevers
IBLemodificationcontrollingproteinactivity.Theamino-terminaltailsofcorehistonesundergolysinemethylationinmultiplesites,termedas“histonecode”or“epigeneticcode”.Lysinemethylationincorehistonesisamajordeterminantfortheformationofactiveandinactiveregionsofthegenomeandthereforeplaysvitalrolesinmultiplecellularevents.Inmostspecies,lysinemethylationoccursprimarilyonhistonesH3(Lys4,9,27,36,79)andH4(Lys20)andhasbeenimplicatedinbothtranscriptionalactivationandsilencing.Methylationinhistonesmodulatedbyspecifichistonemethylases(HMTs)andhistonedemethylases(HDMs)isimpairedinthepathologiesofcancerandotherdiseasesandtherefore,enzymesregulatinghistonelysinemethylationhavebecomepromisingtargetsforanti-cancerdrugs.
Storage&StABIlityTheantibodyisdeliveredinlyophilizedpowder.Uponreceipt,pleasecentrifugetheantibodyat12,000xgfor20seconds,andthenreconstituteitwiththeantibodystabilizerprovided.ThestabilizercontainsPBS,50%glycerol,and0.01%sodiumazide.Pleasestoretheantibodyat-20°C.Avoidrepeatedfreeze/thawcycles.Antibodyisstablefor12monthsfromdateofreconstitution.Equilibratetheantibodytoroomtemperaturefor2minutesandmixbygentlypipettingbeforeuse.**Researchpurposesonly.Theproductisnotintendedtobeusedfortherapeuticordiagnosticpurposesinhumansoranimals.
