Host/IsotypeMouse/IgG1-kappa Clone11G2 SpeciesReactivityBroadReactivity ValidationDataWhitePaperAntibodypedia | Anti-SUMO-2/3antibodyisamousemonoclonalantibody.Theantibodywasraisedagainstfull-lengthrecombinantSUMO-2protein(Uniprot:P61956)combinedwithaproprietarymixofpeptidesthatincludeCQIRFRFDGQPINE.TheantibodyhasbeenshowntoimmunoprecipitateawiderangeofSUMO-2/3targetedproteinsinaHeLacelllysate(Fig.1A).AlinearepitopehasnotbeenidentifiedandASM24appearstorecognizeaconformationalepitope.EachLotofantibodyisqualitycontrolledtoprovideahighbatchtobatchconsistency.TheLotspecificµgpertubecanbefoundintheLotspecificCOAdocumentsatwww.Cytoskeleton.com.ASM24ispurifiedbyProteinGaffinitychromatographyandissuppliedasalyophilizedwhitepowder. EachLotofantibodyisqualitycontrolledtoprovideahighbatchtobatchconsistency.TheLotspecificµgpertubecanbefoundintheLotspecificCOAdocuments.
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ValidatedApplications
Figure1:ImmunoprecipitationusingSUMO-2/3Antibody DenaturedcelllysateswerepreparedfromHS43,CT37andKDS210(HS43:HeatShocktreated(43°Cfor10min),CT37:untreatedandKDS2:shRNASUMO-2knockdown).1mgoflysatewasusedfortheimmunoprecipitationofSUMO-2/3conjugates.IPexperimentswereperformedbytheprotocolpresentedinIPandWBMethod.Westernblotsofimmunopre-cipitatedproteinsweredevelopedusinganti-SUMO-2/3(Cytoskeleoncat#ASM23)(A)oranti-TFII-Iantibody(B).(A)Star(*)andcircle(o)indicateheavyandlightchainsofantibodies.Un-conjugatedfreeSUMOisdenotedbyatriangle.(B)UnconjugatedTFII-IisvisIBLenear120kDa.MultiplebandsindicatethatTFII-IisSUMOylatedbyseveralSUMO-2/3proteins.TFII-IhaspreviouslybeenreportedtobeatargetforSumoylation10,11.ToseethefullImmunoprecipitationprotocol,seetheproductdatasheet. |  |
Figure2:ImmunofluorescenceofHeLacellsinmetaphasewithSUMO-2/3Antibody HeLacellswerestainedandvisualizedbywidefieldfluorescencemicroscopyasde-scribedintheIFmethodbelow.Thecellswerestainedagainstα/β-tubulin(sheepanti-tubulinAb,Cat#ATN02,green)andSUMO-2/3(11G2,red).DNAwasstainedwithDAPI.MitoticcellsinmetaphasewereimagedwithaZeissAxioObserver.Z1microscope(1.4NA63Xobjective).ThelocalizationofSUMO2/3-conjugatesatchromosomescanbeobservedduringmitosisashasbeenpreviouslyreported12.ToseethefullImmunofluorescenceprotocol,seetheproductdatasheet. |  |
Figure3:ImmunofluorescenceofHeLacellsininterphasewithSUMO-2/3Antibody HeLacellswerestainedandvisualizedbywidefieldfluorescencemicroscopyasde-scribedintheIFmethodbelow.Thecellswerestainedagainstβ-tubulin(sheepanti-tubulinAb,Cat#ATN02,green)andSUMO-2/3(11G2,red).DNAwasstainedwithDAPI.CellsininterphasewereimagedwithaZeissAxioObserver.Z1microscope(1.4NA63Xobjective).PMLnuclearbodies(nucleardots)werevisibleinSUMO-2/3stainingashasbeenpreviouslyreported5.ToseethefullImmunofluorescenceprotocol,seetheproductdatasheet. |  |
Formoreinformationcontact:signalseeker@cytoskeleton.com
AssociatedProducts:
Signal-Seeker™SUMOylation2/3DetectionKit(Cat.#BK162)
Signal-Seeker™SUMOylation2/3AffinityBeads(Cat.#ASM24-beads)
Signal-Seeker™:BlastR™RapidLysatePrepKit(Cat.#BLR01)
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