Cat#:AAR-019LyophilizedPowderLyophilizedPowderThisproductisfreezedried.Allwatermoleculeshavebeenremoved.AntigenIncl.AntigenIncl.ThisantibodyisshippedwithitsantigenFREEofcharge!ShippedatRoomTemp.ShippedatRoomTemp.Thislyophilizedproductisshippedatroomtemperature.Pleaseseeitscertificateofanalysisforfurtherstorageinstructions.Type:PolyclonalSource:RabbitReactivity:h,m,rImmunogenPeptide(C)EPVPPDERF*SGITEE,correspondingtoaminoacidresidues231-245ofratα1D-adrenoceptorwithreplacementofcysteine240(C240)withserine(*S)(AccessionP23944).3rdextracellularloop.Accession(Uniprot)NumberP23944GeneID29413PeptideconfirmationConfirmedbyaminoacidanalysisandmassspectrometry.HomologyRat,human,mouse-14/15aminoacidresiduesidentical.PurityAffinitypurifiedonimmobilizedantigen.FormulationLyophilizedpowder.Reconstitutedanitbodycontainsphosphatebufferedsaline(PBS),pH7.4,1%BSA,0.05%NaN3.StoragebeforereconstitutionTheantibodyshipsasalyophilizedpowderatroomtemperature.Uponarrival,itshouldbestoredat-20°C.Reconstitution50μlor0.2mldoubledistilledwater(DDW),dependingonthesamplesize.Antibodyconcentrationafterreconstitution0.8mg/ml.StorageafterreconstitutionThereconstitutedsolutioncanbestoredat4°Cforupto1week.Forlongerperiods,smallaliquotsshouldbestoredat-20°C.Avoidmultiplefreezingandthawing.Centrifugeallantibodypreparationsbeforeuse(10000xg5min).ControlantigenstoragebeforereconstitutionLyophilizedpowdercanbestoredintactatroomtemperaturefor2weeks.Forlongerperiods,itshouldbestoredat-20°C.Controlantigenreconstitution100µldoubledistilledwater(DDW).Controlantigenstorageafterreconstitution-20°C.PreadsorptionControl1μgpeptideper1μgantibody.StandardqualitycontrolofeachlotWesternblotanalysis.Applications:ic,ifc,ih,lci,wbMayalsoworkin:ipWesternblotWesternblotanalysisofratbrainstem(lanes1and3),mousebrain(lanes2and4),SH-SY5Y(lanes5and7)andJurkat(lanes6and8)lysates:1,2,5,6.Anti-α1D-AdrenergicReceptor(extracellular) Antibody(#AAR-019),(1:200).3,4,7,8.Anti-α1D-AdrenergicReceptor(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.ImmunohistochemistryExpressionofα1D-AdrenoceptorinratheartImmunohistochemicalstainingofratheartparaffinembeddedsectionsusingAnti-α1D-AdrenergicReceptor(extracellular)Antibody(#AAR-019),(1:100).α1D-Adrenoceptorisexpressedincardiomyocytesofthemyocardiumandinthesmoothmuscleofthebloodvessels(arrows).Hematoxilinisusedasthecounterstain.Expressionofα1D-AdrenoceptorinratneocortexImmunohistochemicalstainingofα1D-adrenoceptorinratneocortexusingAnti-α1D-AdrenergicReceptor(extracellular)Antibody(#AAR-019). A.Mostintensestainingofα1D-adrenoceptor(green)appearsinapicaldendrites(thinhorizontalarrows)butalsointhesoma(thickhorizontalarrow).Fewcorticalinterneuronsexpressα1D-adrenoceptor.B.Thesamesectionwasalsostainedforparvalbuminandonecell(markedwithaverticalarrow)alsoexpressesα1D-adrenoceptor.IndirectflowcytometryIndirectflowcytometryanalysisofintactliveJurkatcells:___Unstainedcells.___Cells+Anti-α1D-AdrenergicReceptor(extracellular) Antibody(#AAR-019),(5-10µgantibody/0.5x106cells).Thecontrolantigenisnotsuitableforthisapplication.Livecellimaging/ImmunocytochemistryExpressionofα1D-Adrenoceptor inratPC12cellsCellsurfacedetectionof α1D-AdrenoceptorinintactlivingratpheochromocytomaPC12cells.A.ExtracellularstainingofcellsusingAnti-α1D-AdrenergicReceptor (extracellular) Antibody(#AAR-019)(1:100) followedbygoatanti-rabbit-AlexaFluor-594secondaryantibody.B.MergeofAandliveviewofthecells.ReferencesIUPHARRECEPTORDATABASE|ADRENOCEPTORSPiascik,M.T.andPerez,D.M.(2001)J.Pharmacol.Exp.Ther.298,403.Chen,Z.J.andMinneman,K.P.(2005)ActaPharmacol.Sin.26,1281.Tanoue,A.etal.(2002)J.Clin.Invest.109,765.Chen,Q.etal.(2005)J.Urol.174,370.ScientificbackgroundAdrenergicreceptors(alsocalledadrenoceptors)arethereceptorsforthecatecholaminesadrenalineandnoradrenaline(calledepinephrineandnorepinephrineintheUnitedStates).Adrenalineandnoradrenalineplayimportantrolesinthecontrolofbloodpressure,myocardialcontractilerateandforce,airwayreactivity,andavarietyofmetabolicandcentralnervoussystemfunctions. AdrenergicreceptorsaremembersoftheG-proteincoupledreceptor(GPCR)superfamilyofmembraneproteins.Theyshareacommonstructureofsevenputativetransmembranedomains,anextracellularaminoterminus,andacytoplasmiccarboxylterminus.Adrenoceptorsaredividedintothreetypes:α1,α2 andβ-adrenoceptors.Eachtypeisfurtherdividedintoatleastthreesubtypes: α1A, α1B,α1D, α2A, α2B, α2C,β1, β2, β31,2.Theadrenoceptors areexpressedinnearlyallperipheraltissuesandinthecentralnervoussystem1,2. α1-Adrenergicreceptorsplayanimportantroleinthephysiologicalresponsetoepinephrineandnorepinephrine,particularlyinthecardiovascularsystem.Allthreeclonedα1 receptors(α1A,α1B,andα1D)coupletoGq/11.Whiletheircellulardistributionisclear,theirfunctionalroleislesssoduetoalackofspecificagoNISTs/antagonists3. Asthesereceptorslackspecificagonists/antagonists,knock-out(KO)studiesinmicehaveshedsignificantlightontheircellularrole.Individualα1-adrenoceptorKOmicedonotdisplayphysicalabnormalitiesingeneral.SimilarKOstudiesindicatethatallα1 subtypesseemtobeinvolvedintheregulationofbloodpressure3.Also,studiesindicatethatα1D-adrenoceptorseemstoplayanimportantroleinaorticcontraction4,renalfunction,nociception,basallocomotoractivity3 aswellasbladdercontraction5. 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-RatLyophilizedPowderControlAntigenIncludedClickheretoreceivea50µlfreetrialsample!CertificateofAnalysisMSDSLastupdate:19/03/2019AlomoneLabsispleasedtoofferahighlyspecificantibodydirectedagainstanextracellularepitopeoftheratα1D-adrenergicreceptor.Anti-α1D-AdrenergicReceptor(extracellular)Antibody(#AAR-019)canbeusedinwesternblot,indirectflowcytometryandimmunohistochemicalapplications.Ithasbeendesignedtorecognizeα1D-adrenoceptorfrommouse,ratandhumansamples.Forresearchpurposesonly,notforhumanuseWesternblotWesternblotanalysisofratbrainstem(lanes1and3),mousebrain(lanes2and4),SH-SY5Y(lanes5and7)andJurkat(lanes6and8)lysates:1,2,5,6.Anti-α1D-AdrenergicReceptor(extracellular) Antibody(#AAR-019),(1:200).3,4,7,8.Anti-α1D-AdrenergicReceptor(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.

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