Host/IsotypeMouse/IgG2b Clone27B10.4 SpeciesReactivityAllspecies ValidationDataWhitePaperCustomerTestmimonials | Anti-phosphotyrosineantibodyisatyrosinephosphorylationspecificantibodythatispartoftheSignal-Seeker™productline.APY03anti-phosphotyrosinemousemonoclonalantibodyclone27B10hasbeentestedinwesternblot,immunofluorescence,immunoprecipitationandELISAwithexcellentspecificityandlowbackgroundwithstrongpotentsignals.APY03phosphotyrosineantibodyrecognizesawiderangeoftyrosinephosphorylatedproteinsincellstreatedwithkinaseinhibitors,hydrogenperoxide(H2O2)andsodiumvanadate(Figure1). APY03isamousemonoclonalantibody27B10thatrecognizesphosphorytatedtyrosineresidues.APY03wasraisedagainstaproprietarymixtureofphosphotyrosinepeptidesconjugatedtoKLH.APY03candetect10ngofphosphotyrosine-labeledbovineserumalbumin(seeCertificateofAnalysis[COA]).APY03ispurifiedbyproteinGaffinitychromatographyandissuppliedasalyophilizedwhitepowder.EachLotofantibodyisqualitycontrolledtoprovideahighbatchtobatchconsistency.TheLotspecificµgpertubecanbefoundintheLotspecificCOAdocumentsatwww.Cytoskeleton.com.APY03showshighspecificitytophosphotyrosinepeptidesanddoesnotcross-reactwithphosphoserine/threoninepeptidesinawesternblotoranELISAassay(Figures1&4).
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ValidatedApplications
Figure1:WesternBlotusingPhosphotyrosineAntibody APY03ishighlyspecificforp-Tyranddoesnotrecognizep-Serorp-Thrtaggedproteins.ToseethefullWesternblotprotocol,seetheproductdatasheet. |
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Figure2:ImmunoprecipitationusingPhosphotyrosineAntibody APY03wasabletoenrichawiderangeoftyrosine-phosphorylatedproteinsfromNIH3T3cellstreatedwithH2O2-activatedorthovanadate.NosignalwasdetectedwithProteinGbeadcontrolwithoutAPY03.ToseethefullImmunoprecipitationprotocol,seetheproductdatasheet. |  |
Figure3:ImmunofluorescenceusingPhosphotyrosineAntibody HumanepidermoidcarcinomaA431cells,untreated(3A)ortreated(3B)withEGF(100ng/mlfor3minutes),andNIH3T3,untreated(3C)ortreated(3D)withH2O2-activatedsodiumorthovanadate(100µMfor10minutes),werestainedasdescribedinthemethod.PhosphotyrosineandnucleiwerevisualizedingreenfluorescenceandblueDAPIstaining,respectively.ToseethefullImmunofluorescenceprotocol,seetheproductdatasheet. |  |
Figure4:ELISAusingPhosphotyrosineAntibody DatashowthatAPY03doesnotcrossreactwithphosphothreonineandphoophoserinemodifiedBSA.ToseethefullELISAprotocol,seetheproductdatasheet. |  |
Formoreinformationcontact:signalseeker@cytoskeleton.com
AssociatedProducts:
Signal-Seeker™PhosphotyrosineDetectionKit(Cat.#BK160)
Signal-Seeker™PhosphotyrosineAffinityBeads(Cat.#APY03-beads)
Signal-Seeker™:BlastR™RapidLysatePrepKit(Cat.#BLR01)
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