Cat#:APZ-031LyophilizedPowderLyophilizedPowderThisproductisfreezedried.Allwatermoleculeshavebeenremoved.AntigenIncl.AntigenIncl.ThisantibodyisshippedwithitsantigenFREEofcharge!ShippedatRoomTemp.ShippedatRoomTemp.Thislyophilizedproductisshippedatroomtemperature.Pleaseseeitscertificateofanalysisforfurtherstorageinstructions.Type:PolyclonalSource:RabbitReactivity:m,rImmunogenPeptide(C)RGAKRSRPSKLTED,correspondingtoaminoacidresidues128-141ofratAKAP150(Accession P24587).Intracellular.Accession(Uniprot)NumberP24587GeneIDP24587PeptideconfirmationConfirmedbyaminoacidanalysisandmassspectrometry.HomologyMouse–13/14aminoacidresiduesidentical.PurityAffinitypurifiedonimmobilizedantigen.FormulationLyophilizedpowder.Reconstitutedantibodycontainsphosphatebufferedsaline(PBS),pH7.4,1%BSA,0.05%NaN3.SpecificityTheantibodywon’trecognizethehumanAKAP150homologue(alsoknownasAKAP79).StoragebeforereconstitutionTheantibodyshipsasalyophilizedpowderatroomtemperature.Uponarrival,itshouldbestoredat-20°C.Reconstitution25µl,50µlor0.2mldoubledistilledwater(DDW),dependingonthesamplesize.Antibodyconcentrationafterreconstitution0.8mg/ml.StorageafterreconstitutionThereconstitutedsolutioncanbestoredat4°Cforupto1week.Forlongerperiods,smallaliquotsshouldbestoredat-20°C.Avoidmultiplefreezingandthawing.Centrifugeallantibodypreparationsbeforeuse(10000xg5min).ControlantigenstoragebeforereconstitutionLyophilizedpowdercanbestoredintactatroomtemperaturefor2weeks.Forlongerperiods,itshouldbestoredat-20°C.Controlantigenreconstitution100µldoubledistilledwater(DDW).Controlantigenstorageafterreconstitution-20ºC.PreadsorptionControl1µgpeptideper1µgantibody.StandardqualitycontrolofeachlotWesternblotanalysis.Applications:wbMayalsoworkin:ic,ifc,ih,ipWesternblotWesternblotanalysisofratbrainmembranes(lanes1and5),mousebrainmembranes(lanes2and6),ratdorsalrootganglionlysate(lanes3and7)andmouselunglysates(lanes4and8):1-4. Anti-AKAP150Antibody(#APZ-031),(1:200).5-8.Anti-AKAP150Antibody,preincubatedwiththecontrolpeptideantigen.ReferencesSchwartz,J.H.(2001)Proc.Natl.Acad.Sci.USA.98,13482.Feliciello,A.etal.(2001)J.Mol.Biol.308,99.Adler,E.M.(2014)J.Gen.Physiol.143,313.Esseltine,J.L.andScott,J.D.(2013)Trends.Pharmacol.Sci.34,648.Ting,J.T.etal.(2012)Annu.Rev.Neurosci.35,49.Nieves-Cintron,M.etal.(2015)Cell.Signal.Onlinepublication.ScientificbackgroundA-Kinaseanchorproteins(AKAPs)areagroupofstructurallydiverseproteinsthatbindtotheregulatoryunitofProteinKinaseA(PKA)todiscretelocationswithinthecell1.AKAPsincludetwoconservedstructuralmodules:AtargetingdomainthatservesasascaffoldandmembraneanchorthatconsistsoftwoNH2-terminalstructuralmodulesthatarehighlyconservedamongmammalianhomologues.TheyalsohaveatetheringdomainthatinteractswithPKAregulatorysubunitswhichconsistsofashortamphipathichelix.AlternativesplicingcanshuffletargetingandtetheringdomainstogenerateavarietyofAKAPswithdifferenttargetingspecificity.AlthoughAKAPshavebeenidentifiedonthebasisoftheirinteractionwithPKA,theyalsobindothersignalingmolecules,mainlyphosphatasesandkinasesthatregulateAKAPtargetingandactivateothersignaltransductionpathways2.SometypesofAKAPsinteractwith CaV1.1 and CaV1.2.ThesevoltagegatedcalciumchannelsarephosphorylatedbyPKAafterβ-adrenergicactivation.ThisphosphorylationisenabledbythebindingofAKAP150tothedistalC-terminalofCaV1.1andCaV1.2.BindingofdifferentAKAPsthatinteractwithdistinctsignalingpartnerstothecalciumchannelintracellularC-terminaldomainmayacttofine-tunechannelactivityanddownstreamcalcium-mediatedprocesses3.AsinglesubtypeofneuronalAKAPisexpressedwidelyinthebrainandishighlyconservedinmouse(150kDA),bovine(75kDA)andhuman(79kDA).APAK150targetsPKAIItothepostsynapticCytoskeletonandperikaryaofneurons.ItisexpressedinPurkinjecellsandinolfactorybulbneurons,basalganglia,cerebralcortexandotherforebrainareas.AKAP150isconcentratedinprimarybranchesofdendritesinassociationwithmicrotubules.ThelocalizationofPKAIIβinbrainoverlapswiththatofAKAP150,suggestingthatAKAP150istherelevantPKAIIβanchorproteininthebrain2.InhibitionofeitherAKAP150or TRPV1 bindingsitescausesanabolishmentofPKC-dependentTRPV1sensitizationtocapsaicinstimulithusmakingtheAKAP150-TRPV1interactionaviabletargetfornovelanalgesics4.AKAP150interactswithanothergroupofscaffoldingproteinsknownastheSAP90/PSD95-associatedproteinfamily.AKAP150interactsdirectlywiththeSH3andGKdomainsof PSD95 and SAP97.Theseproteinshavespecificrolesinsynapticlocalizationof NMDARs and AMPARs respectively5.Postmyocardialinfarction,AKAP150-mediatedtargetingofcalcineurintosarcolemmalmicro-domainsinventricularmyocytescontributestothelocalandacutegeneremodelingeventsthatleadtothedown-regulationof KV currents6.Applicationkey:CBE-Cell-basedELISA,FC-Flowcytometry,ICC-Immunocytochemistry,IE-IndirectELISA,IFC-Indirectflowcytometry,IHC-Immunohistochemistry,IP-Immunoprecipitation,LCI-Livecellimaging,N-Neutralization,WB-WesternblotSpeciesreactivitykey:H-Human,M-Mouse,R-RatLyophilizedPowderControlAntigenIncludedClickheretoreceivea25µlfreetrialsample!CertificateofAnalysisMSDSLastupdate:06/03/2019Anti-AKAP150Antibody(#APZ-031)isahighlyspecificantibodydirectedagainstanepitopeoftheratprotein.Theantibodycanbeusedinwesternblotanalysis.IthasbeendesignedtorecognizeAKAP150frommouseandratsamples.TheantibodywillunlikelyrecognizeAKAP150fromhumansamples.Forresearchpurposesonly,notforhumanuseWesternblotWesternblotanalysisofratbrainmembranes(lanes1and5),mousebrainmembranes(lanes2and6),ratdorsalrootganglionlysate(lanes3and7)andmouselunglysates(lanes4and8):1-4. Anti-AKAP150Antibody(#APZ-031),(1:200).5-8.Anti-AKAP150Antibody,preincubatedwiththecontrolpeptideantigen.

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