LyophilizedPowderLyophilizedPowder
Thisproductisfreezedried.Allwatermoleculeshavebeenremoved.
AntigenIncl.AntigenIncl.
ThisantibodyisshippedwithitsantigenFREEofcharge!
ShippedatRoomTemp.ShippedatRoomTemp.
Thislyophilizedproductisshippedatroomtemperature.Pleaseseeitscertificateofanalysisforfurtherstorageinstructions.
Type:Polyclonal
Source:Rabbit
Reactivity:m,r
ImmunogenPeptide(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
WesternblotWesternblotanalysisofratbrainmembranes(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.
LyophilizedPowderControlAntigenIncluded
Clickheretoreceivea25µlfreetrialsample!
Lastupdate:06/03/2019
Anti-AKAP150Antibody(#APZ-031)isahighlyspecificantibodydirectedagainstanepitopeoftheratprotein.Theantibodycanbeusedinwesternblotanalysis.IthasbeendesignedtorecognizeAKAP150frommouseandratsamples.TheantibodywillunlikelyrecognizeAKAP150fromhumansamples.
Forresearchpurposesonly,notforhumanuse
WesternblotWesternblotanalysisofratbrainmembranes(lanes1and5),mousebrainmembranes(lanes2and6),ratdorsalrootganglionlysate(lanes3and7)andmouselunglysates(lanes4and8):
1-4. Anti-AKAP150Antibody(#APZ-031),(1:200).
5-8.Anti-AKAP150Antibody,preincubatedwiththecontrolpeptideantigen.