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Millipore/15-103 | PI3 Kinase/Akt/GSK3 Pathway Explorer Minipack/15-103/3 vials/Pk
  • Millipore/15-103 | PI3 Kinase/Akt/GSK3 Pathway Explorer Minipack/15-103/3 vials/Pk

Millipore/15-103 | PI3 Kinase/Akt/GSK3 Pathway Explorer Minipack/15-103/3 vials/Pk

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    • Description
      CatalogueNumber15-103
      BrandFamilyUpstate
      TradeName
      • Upstate
      DescriptionPI3Kinase/Akt/GSK3PathwayExplorerMinipack
      AlternateNames
      • ProteinkinaseB
      • RAC-alphaserine/threonine-proteinkinase
      • racproteinkinasealpha
      • v-aktmurinethymomaviraloncogenehomolog1
      • murinethymomaviral(v-akt)oncogenehomolog-1
      • glycogensynthasekinase3alpha
      • glycogensynthasekinase3beta
      BackgroundInformationPathwayExplorerAntibodyMiniPack:
      EachPathwayExplorerAntibodyMinipackcontainsthreerelatedantibodiesaspartofasignalingcascadeoracombinationoftotalandphosphorylatedformsofkeysignalingtargets.Eachofthethreeantibodiesare30%theoriginalpacksize.FullsizeversionsofeachofthePathwayExplorerantibodiesareavailableforsaleindividuallyunderthesamecatalognumberwiththeremovalof“SP”offofeachone(e.g.05-591SPcanbeorderedas05-591).

      PI3Kinase:
      PI3Kinase(Phosphatidylinositol3-Kinase)isresponsIBLeforphosphorylationofthe3positionoftheinositolringofPI(4,5)P2,togeneratePI(3,4,5)P3,apotentsecondmessengerrequiredforsurvivalsignaling,andinsulinaction.PI3Kinaseisaheterodimericcomplexcomposedofan85kDaregulatorysubunitanda110kDacatalyticsubunit.Tyrosinephosphorylationofgrowthfactorreceptorscreatesdockingsitesforbindingofp85(throughitsSH2domains)onthereceptors;p85bringswithitp110,whichisthenproximaltoitsphospho-lipidsubstrateonthemembrane.PI3KinaseisalsoactivatedbyRas,andbytheβ-γsubunitsofheterotrimericG-proteins.PI3Kinaseisinhibitablebywortmannin,ausefultoolforthestudyofthePI3Kinasesignalingpathway.

      Akt/PKB,PHDomain:
      Akt(proteinkinaseB),aserine/threoninekinase,hasemergedasacriticalenzymeinsignaltransductionpathwaysinvolvedincellproliferation,apoptosis,angiogenesis,anddiabetes.InmammalsthreeisoformsofAkt(α,β,γorAkt1,2,3)arereportedthatexhibitahighdegreeofhomology,butdifferslightlyinthelocalizationoftheirregulatoryphosphorylationsites.Aktαisthepredominantisoforminmosttissues,whereasthehighestexpressionofAktβisobservedintheinsulin-responsivetissues,andAktγisabundantinbraintissue.EachAktisoformiscomposedofthreefunctionallydistinctregions:anN-terminalpleckstrinhomology(PH)domainthatprovidesalipid-bindingmoduletodirectAkttoPIP3,acentralcatalyticdomain,andaC-terminalhydrophobicmotif.

      GSK3α/β:
      Glycogensynthasekinase-3(GSK-3),amultifunctionalserine/threoninekinase,isakeyregulatorofnumeroussignalingpathways.TwoisoformsofGSK-3arereportedinmammals:a51kDaGSK-3αanda47kDaGSK-3β.Thesetwoisoformsexhibitabout98%homologyintheirkinasedomains,butshareonlyabout36%identityinthelast76C-terminalaminoacidresidues.GSK-3αcontainsaglycine-richextensionatitsN-terminus.Aminor(~15%oftotal)splicevariantofGSK-3β,GSK-3β2,hasalsobeenidentified,whichcontainsa13-residueinsertwithinthekinasedomain.ItexhibitsreducedkinaseactivitytowardstheTauproteincomparedwith‘unspliced’GSK-3β.GSK-3β2islocalizedprimarilytoneuronalcellbodies,unlikeunsplicedGSK-3βthatisalsofoundinneuronalprocesses.GSK-3βplaysakeyinhibitoryroleintheWntsignalingpathway.Wntgenesencodealargefamilyofsecreted,cysteine-richproteinsthatareimportantindevelopmentandinmaintenanceofadulttissues.AbnormalitiesinWntsignalingarereportedtopromotebothhumandegenerativediseasesandcancer.

      *Seefullsizeversionsforcorrespondingreferences.
      ProductInformation
      Components
      • 06-195SPAnti-PI3Kinase,p85
      • 05-591SPAnti-Akt/PKB,PHDomain,cloneSKB1
      • 05-412SPAnti-GSK3α/β,clone4G-1E
      Control
      • 05-412SP:IncludedPositiveAntigenControl:Catalog#12-303,Jurkatlysate.Add2.5μLof2-mercaptoethanolper100μLoflysateandboilfor5minutestoreducethepreparation.Use20μgperlaneforminigels.
      • 06-195SP:Jurkatcelllysate.
        IncludedPositiveAntigenControl:Catalog#12-303,Jurkatcelllysate.Add2.5μLof2-mercaptoethanolper100μLoflysateandboilfor5minutestoreducethepreparation.Load20μgofreducedlysateperlaneforminigels.
      Presentation3individualtubescontainingeitherAnti-PI3Kinase,p85;Anti-Akt/PKB,PHDomain,cloneSKB1;orAnti-GSK3α/β,clone4G-1E
      PropertiesEachvialis30%thesizeoftheparentcatalognumber
      StorageandShippingInformation
      Applications
      ApplicationThisAntibodypackcontainsAnti-PI3KinaseAntibody,Anti-Akt/PKBAntibody,Anti-GSK3α/βAntibodyvalidatedforuseinEnzymeAssay,ICC,IF,Flow,IP&WB.
      KeyApplications
      • EnzymeAssays
      • Immunocytochemistry
      • Immunofluorescence
      • FlowCytometry
      • Immunoprecipitation
      • WesternBlotting
      ApplicationsNotRecommended
      • Immunoprecipitation
      ApplicationNotes05-412SP:ImmunofluorescenceStaining:
      L6cellsweregrown,fixed,andpermeABIlized.;
      FlowCytometry:
      NCI/H292cellswerestainedwithAnti-GSK3α/β,clone4G-1EconjugatedtoAlexaFluor?555;Immunoprecipitation:Notrecommendedfor05-412SP.

      06-195SP:Immunoprecipitation:5μLofapreviouslotimmunoprecipitatedPI3kinasefromhumanJurkatcelllysateasconfirmedbysubsequentwesternblot.Theantibodywillprecipitatethe85kDasubunit.

      05-591SP:ImmunofluroescenceAnalysis:
      A431cellswerefixed,permeabilized,andstainedwithanti-Akt,cloneSKB1(2μg/mL)conjugatedtoAlexaFluor?488;
      FlowCytometry:NIH/3T3cells(detachedusingtrypsin-EDTA)persamplewerestainedwith0.5μgofanti-Akt/PKB,PHDomain,cloneSKB1(filledhistogram)or1μgofmouseIgG1(openhistogram)followedbyagoatanti-mouseIgG-PEconjugate.

      ImmunoprecipitationKinase:4μgofapreviouslotimmunoprecipitatedactiveAktfrom700μgofL6cellsstimulatedwith15nMIGF-1for2-5minutes.

      ImmunoprecipitationKinaseAssayData:
      ThekinaseactivityofimmunoprecipitatedAkt/PKBwasdeterminedasdescribedaboveusingAkt/SGKSubstratePeptideasasubstrate.Activekinasewasdetectedbycomparingtheincorporationof[32P]intoAkt/SGKSubstratePeptideascatalyzedbyimmunoprecipitates(1mg)ofL6celllysatesstimulatedwith15nMIGF-1.Thetestresultsareshowninthetabletotheleft.

      BIOLOGicalInformation
      Immunogen05-412SP:18residuesyntheticpeptide(C-KQLLHGEPNVSYICSRY)correspondingtoaminoacids203-219derivedfromaregionofthecatalyticdomain(betweensuBDomainVIIandVIII)oftheDrosophilaGSK3/shaggyenzyme,coupledtoovalbumin.Clone4G-1E.;05-591SP:GST-fusionproteincorrespondingtoresidues1-149ofhumanAkt1.;06-195SP:GSTfusionproteinscontaining1)theN-terminalSH2domainofratPI3kinaseand2)thefulllength85kDasubunitofratPI3kinase.
      EpitopeVaries.SeeindividualDataSheets
      Clone05-412SP:4G-1E;05-591SP:SKB1
      ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
      SpeciesReactivity
      • Human
      • Mouse
      • Rat
      • Vertebrates
      • Monkey
      • Yeast(S.pombe)
      • Dictyostelium
      SpeciesReactivityNote05-412SP:Human,Mouse,Rat,S.pombe,andDictyostelium.Becauseoftheextremelyhighconservationofthisimmunogenamongtheknownenzymesofmanydifferentspecies,thisantibodyispredictedtocross-reactwithnearlyallspecieshomologues.

      05-591SP:Human,mouseandrat;otherspeciescross-reactivityunknown.
      EntrezGeneNumber
      EntrezGeneSummaryAKT1:Theserine-threonineproteinkinaseencodedbytheAKT1geneiscatalyticallyinactiveinserum-starvedprimaryandimmortalizedfibroblasts.AKT1andtherelatedAKT2areactivatedbyplatelet-derivedgrowthfactor.Theactivationisrapidandspecific,anditisabrogatedbymutationsinthepleckstrinhomologydomainofAKT1.Itwasshownthattheactivationoccursthroughphosphatidylinositol3-kinase.InthedevelopingnervoussystemAKTisacriticalmediatorofgrowthfactor-inducedneuronalsurvival.Survivalfactorscansuppressapoptosisinatranscription-independentmannerbyactivatingtheserine/threoninekinaseAKT1,whichthenphosphorylatesandinactivatescomponentsoftheapoptoticmachinery.Multiplealternativelysplicedtranscriptvariantshavebeenfoundforthisgene.[providedbyRefSeq].

      P13KINASE:Phosphatidylinositol3-kinasephosphorylatestheinositolringofphosphatidylinositolatthe3-primeposition.Theenzymecomprisesa110kDcatalyticsubunitandaregulatorysubunitofeither85,55,or50kD.Thisgeneencodesthe85kDregulatorysubunit.Phosphatidylinositol3-kinaseplaysanimportantroleinthemetabolicactionsofinsulin,andamutationinthisgenehasbeenassociatedwithinsulinresistance.Alternativesplicingofthisgeneresultsinthreetranscriptvariantsencodingdifferentisoforms.[providedbyRefSeq].

      GeneSymbol
      • AKT
      • Akt1
      • GSK3
      • GlycogenSynthaseKinase3
      • PKB
      • PKB-ALPHA
      • PRKBA
      • PI3Kinase
      • PI3K
      • phosphatidylinositol3-kinaseregulatorysubunit
      • phosphatidylinositol3kinase
      • PIK3R1
      UniProtNumber
      UniProtSummaryPI3Kinase,p85:
      FUNCTION:Bindstoactivated(phosphorylated)protein-Tyrkinases,throughitsSH2domain,andactsasanadapter,mediatingtheassociationofthep110catalyticunittotheplasmamembrane.Necessaryfortheinsulin-stimulatedincreaseinglucoseuptakeandglycogensynthesisininsulin-sensitivetissues.
      Subunitstructure:Heterodimerofap110(catalytic)andap85(regulatory)subunits.InteractswithphosphorylatedTOM1L1.InteractswithphosphorylatedLIME1uponTCRand/orBCRactivation.InteractswithSOCS7.InteractswithRUFY3Bysimilarity.InteractswithphosphorylatedLAT,LAX1andTRAT1uponTCRactivation.InteractswithCBLB.InteractswithHIV-1NeftoactivatetheNefassociatedp21-activatedkinase(PAK).ThisinteractiondependsontheC-terminusofbothproteinsandleadstoincreasedproductionofHIV.InteractswithHCVNS5A.TheSH2domainsinteractwiththeYTHMmotifofphosphorylatedINSRinvitro.Alsointeractswithtyrosine-phosphorylatedIGF1Rinvitro.InteractswithCD28andCD3ZuponT-cellactivation.InteractswithIRS1andphosphorylatedIRS4,aswellaswithNISCHandHCST.
      TISSUESPECIFICITY:Isoform2isexpressedinskeletalmuscleandbrain,andatlowerlevelsinkidneyandcardiacmuscle.Isoform2andisoform4arepresentinskeletalmuscle(atproteinlevel).
      DOMAIN:TheSH3domainmediatesthebindingtoCBLB,andtoHIV-1Nef.
      Post-translationalmodification:PolyubiquitinatedinT-cellsbyCBLB;whichdoesnotpromoteproteasomaldegradationbutimpairsassociationwithCD28andCD3ZuponT-cellactivation.

      AKT:
      FUNCTION:Generalproteinkinasecapableofphosphorylatingseveralknownproteins.PhosphorylatesTBC1D4.Signalsdownstreamofphosphatidylinositol3-kinase(PI(3)K)tomediatetheeffectsofvariousgrowthfactorssuchasplatelet-derivedgrowthfactor(PDGF),epidermalgrowthfactor(EGF),insulinandinsulin-likegrowthfactorI(IGF-I).Playsaroleinglucosetransportbymediatinginsulin-inducedtranslocationoftheGLUT4glucosetransportertothecellsurface.MediatestheantiapoptoticeffectsofIGF-I.Mediatesinsulin-stimulatedproteinsynthesis,partlybyplayingaroleinbothinsulin-inducedphosphorylationof4E-BP1andininsulin-inducedactivationofp70S6kinase.Promotesglycogensynthesisbymediatingtheinsulin-inducedactivationofglycogensynthase.
      CATALYTICACTIVITY:ATP+aprotein=ADP+aphosphoprotein.
      ENZYMEREGULATION:Threespecificsites,oneinthekinasedomain(Thr-308)andthetwootheronesintheC-terminalregulatoryregion(Ser-473andTyr-474),needtobephosphorylatedforitsfullactivation.
      SUBUNIT:InteractswithCENTG1isoform2(PIKE-A)inthepresenceofguaninenucleotides.TheC-terminusinteractswithCCDC88A/GRDNandTHEM4.InteractswithAKTIP.Interacts(viaPHdomain)withMTCP1,TCL1AANDTCL1B.InteractswithCDKN1B;theinteractionphosphorylatesCDKN1Bpromoting14-3-3bindingandcell-cycleprogression.
      SUBCELLULARLOCATION:Cytoplasm.Nucleus.Cellmembrane.Note=Nucleusafteractivationbyintegrin-linkedproteinkinase1(ILK1).NucleartranslocationisenhancedbyinteractionwithTCL1A.
      TISSUESPECIFICITY:Inallhumancelltypessofaranalyzed.
      DOMAIN:BindingofthePHdomaintothephosphatidylinositol3-kinasealpha(PI(3)K)resultsinitstargetingtotheplasmamembrane.
      DOMAIN:TheAGC-kinaseC-terminalmediatesinteractionwithTHEM4.
      PTM:PhosphorylationonThr-308,Ser-473andTyr-474isrequiredforfullactivity.Ser-473phosphorylationbytheRictor-mTorcomplexfavorsThr-308phosphorylationbyPDPK1.Ser-473phosphorylationisenhancedbyinteractionwithCENTG1isoform2(PIKE-A).Ser-473phosphorylationisenhancedinfocalcorticaldysplasiaswithTaylor-typeballooncells.

      GSK3:
      FUNCTION:ParticipatesintheWntsignalingpathway.Implicatedinthehormonalcontrolofseveralregulatoryproteinsincludingglycogensynthase,MYBandthetranscriptionfactorJUN.PhosphorylatesJUNatsitesproximaltoitsDNA-bindingdomain,therebyreducingitsaffinityforDNA.PhosphorylatesMUC1inbreastcancercells,anddecreasestheinteractionofMUC1withCTNNB1/beta-catenin.
      CATALYTICACTIVITY:ATP+[tauprotein]=ADP+[tauprotein]phosphate.
      ENZYMEREGULATION:InhibitedwhenphosphorylatedbyAKT1.
      SUBUNIT:MonomerBysimilarity.InteractswithCABYR,MUC1,NINandPRUNE.
      TISSUESPECIFICITY:Expressedintestis,thymus,prostateandovaryandweaklyexpressedinlung,brainandkidney.
      POST-TRANSLATIONALMODIFICATION:PhosphorylatedbyAKT1andILK1.
      SEQUENCESIMILARITIES:Belongstotheproteinkinasesuperfamily.CMGCSer/Thrproteinkinasefamily.GSK-3subfamily.Contains1proteinkinasedomain.
      MolecularWeight05-412SP:51kDa(α)or46kDa(β);05-591SP:60kDa;06-195SP:85kDa
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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