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Alomone/Anti-AKAP150 Antibody/APZ-031/25 µl

Cat#:APZ-031
  • LyophilizedPowder
  • AntigenIncl.
  • ShippedatRoomTemp.
    • Type:Polyclonal
    Source:Rabbit
    Reactivity:m,r
    Immunogen
    Peptide(C)RGAKRSRPSKLTED,correspondingtoaminoacidresidues128-141ofratAKAP150(Accession P24587).Intracellular.
    Accession(Uniprot)NumberP24587
    GeneIDP24587
    PeptideconfirmationConfirmedbyaminoacidanalysisandmassspectrometry.
    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:wb
    Mayalsoworkin:ic,ifc,ih,ip
    Westernblot
    Westernblotanalysisofratbrainmembranes(lanes1and5),mousebrainmembranes(lanes2and6),ratdorsalrootganglionlysate(lanes3and7)andmouselunglysates(lanes4and8):
    1-4. Anti-AKAP150Antibody(#APZ-031),(1:200).
    5-8.Anti-AKAP150Antibody,preincubatedwiththecontrolpeptideantigen.
    References
    1. Schwartz,J.H.(2001)Proc.Natl.Acad.Sci.USA.98,13482.
    2. Feliciello,A.etal.(2001)J.Mol.Biol.308,99.
    3. Adler,E.M.(2014)J.Gen.Physiol.143,313.
    4. Esseltine,J.L.andScott,J.D.(2013)Trends.Pharmacol.Sci.34,648.
    5. Ting,J.T.etal.(2012)Annu.Rev.Neurosci.35,49.
    6. Nieves-Cintron,M.etal.(2015)Cell.Signal.Onlinepublication.
    Scientificbackground

    A-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-Westernblot

    Speciesreactivitykey:

    H-Human,M-Mouse,R-Rat
    LyophilizedPowderControlAntigenIncluded
    receivea25µlfreetrialsample!
    Lastupdate:06/03/2019

    Anti-AKAP150Antibody(#APZ-031)isahighlyspecificantibodydirectedagainstanepitopeoftheratprotein.Theantibodycanbeusedinwesternblotanalysis.IthasbeendesignedtorecognizeAKAP150frommouseandratsamples.TheantibodywillunlikelyrecognizeAKAP150fromhumansamples.

    Forresearchpurposesonly,notforhumanuse
    Westernblot
    Westernblotanalysisofratbrainmembranes(lanes1and5),mousebrainmembranes(lanes2and6),ratdorsalrootganglionlysate(lanes3and7)andmouselunglysates(lanes4and8):
    1-4. Anti-AKAP150Antibody(#APZ-031),(1:200).
    5-8.Anti-AKAP150Antibody,preincubatedwiththecontrolpeptideantigen.
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