Cat#:AAR-003LyophilizedPowderLyophilizedPowderThisproductisfreezedried.Allwatermoleculeshavebeenremoved.AntigenIncl.AntigenIncl.ThisantibodyisshippedwithitsantigenFREEofcharge!ShippedatRoomTemp.ShippedatRoomTemp.Thislyophilizedproductisshippedatroomtemperature.Pleaseseeitscertificateofanalysisforfurtherstorageinstructions.Type:PolyclonalSource:RabbitReactivity:h,m,rImmunogenPeptideKDSATNN*STEPWDGTTNESC,correspondingtoaminoacidresidues147-166ofhuman A2BARwithreplacementofcysteine154(C154)withserine(*S)(AccessionP29275).2ndextracellularloop.Accession(Uniprot)NumberP29275GeneID136PeptideconfirmationConfirmedbyaminoacidanalysisandmassspectrometry.HomologyRat-14/19aminoacidresiduesidentical;mouse-13/19aminoacidresiduesidentical.PurityAffinitypurifiedonimmobilizedantigen.FormulationLyophilizedpowder.Reconstitutedantibodycontainsphosphatebufferedsaline(PBS),pH7.4,1%BSA,0.05%NaN3.StoragebeforereconstitutionTheantibodyshipsasalyophilizedpowderatroomtemperature.Uponarrival,itshouldbestoredat-20°C.Reconstitution25µl,50µlor0.2mldoubledistilledwater(DDW),dependingonthesamplesize.Antibodyconcentrationafterreconstitution0.75mg/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:ipWesternblotWesternblotanalysisofHL-60(lanes1and2)cellline,ratbrain(lanes3and4)andmousebrain(lanes5and6)lysates:1,3,5.Anti-Adenosine A2B Receptor (extracellular)Antibody(#AAR-003),(1:200).2,4,6.Anti-AdenosineA2B Receptor(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.ImmunohistochemistryExpressionofAdenosine A2B ReceptorinratlungImmunohistochemicalstainingofratlungparaffinembeddedsectionsusingAnti-Adenosine A2B Receptor(extracellular) Antibody(#AAR-003),(1:50).Stainingispresentintherespiratoryepitheliumofthebronchiole(Br)aswellasinthepneumonocytesofthealveolarwall(alveoli,(A).ColorreactionwasobtainedwithSuperPictureHRP-conjugatedpolymer(Zymed)followedbyDAB.Hematoxilinisusedasthecounterstain.IndirectflowcytometryRatenterochromaffin(EC)cells(1:500),(Damen,R. etal. (2013) PLoSONE 8, e62607.).Livecellimaging/ImmunocytochemistryExpressionofAdenosine A2B Receptor inratPC12cellsCellsurfacedetectionof Adenosine A2B ReceptorinintactlivingratpheochromocytomaPC12cells.A.ExtracellularstainingofcellsusingAnti-Adenosine A2B Receptor(extracellular) Antibody(#AAR-003), (1:100)followedbygoatanti-rabbit-AlexaFluor-594secondaryantibody.B.MergeofAandliveviewofthecells.ReferencesOkusa,M.D.(2002)Am.J.Physiol.RenalPhysiol.282,F10.Fredholm,B.B.etal.(2001)Pharmacol.Rev.53,527.Nakata,H.(1989)J.Biol.Chem.264,16545.Baraldi,P.G.etal.(2006)Curr.Med.Chem.13,3467.Polosa,R.(2002)Eur.Respir.J.20,488.ScientificbackgroundAdenosineisanendogenousnucleosidegeneratedlocallyintissuesunderconditionsofhypoxia,ischemia,orinflammation.Itmodulatesavarietyofphysiologicalfunctionsinmanytissuesincludingthebrainandheart.1,2 Adenosineexertsitsactionsviafourspecificadenosinereceptors(alsonamedP1purinergicreceptors): AdenosineA1 Receptor (A1AR), AdenosineA2A Receptor (A2AAR),AdenosineA2B Receptor (A2BAR),and AdenosineA3 Receptor (A3AR).AllareintegralmembraneproteinsandaremembersoftheGprotein-coupledreceptorsuperfamily.Theyshareacommonstructureofsevenputativetransmembranedomains,anextracellularaminoterminus,acytoplasmiccarboxylterminus,andathirdintracellularloopimportantinbindingGproteins.1-3Thevariousadenosinereceptorscanbedistinguishedonthebasisoftheirdistinctmolecularstructures,distincttissuedistributions,anddifferentialselectivityforadenosineanalogs.1-4TheadenosineA2receptorsubtypewassuBDividedaccordingtoaffinityforadenosine;A2AARhashighaffinityandA2BARhaslowaffinity.A2BARisexpressedinmastcells,neutrophils,monocytes,macrophages,dendriticcells,Tcells,bronchialepithelium,intestinalepithelium,smoothmuscle,brain,andothercells.5 Inintestinalepithelia,A2BARwasfoundtobethemajoradenosinereceptorsubtypeexpressedandisthoughttobeinvolvedindiarrhealdiseases.Itisalsobelievedtobeinvolvedinthepathogenesisofchronicairwayinflammatorydiseases.4 Thus,antagoNISTsofA2BARmightbeeffectiveforthetreatmentofinflammatorygastrointestinaltractdisorders4 aswellasasthmaandchronicobstructivepulmonarydisease.5Applicationkey: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:19/03/2019Anti-Adenosine A2B Receptor(extracellular)Antibody(#AAR-003)isahighlyspecificantibodydirectedagainstanextracellularepitopeofhumanadenosineA2B receptor(A2BAR).Theantibody canbeusedinwesternblot,immunohistochemistry,livecellimagingandindirectflowcytometryapplications.IthasbeendesignedtorecognizeA2BAR fromhuman,ratandmousesamples.Forresearchpurposesonly,notforhumanuseWesternblotWesternblotanalysisofHL-60(lanes1and2)cellline,ratbrain(lanes3and4)andmousebrain(lanes5and6)lysates:1,3,5.Anti-Adenosine A2B Receptor (extracellular)Antibody(#AAR-003),(1:200).2,4,6.Anti-AdenosineA2B Receptor(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.

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