Type:GuineaPigIgG
Applications:ICC;IHC
E=ELISA;FACS;FC=FlowCytometry;FPLC=FastProteinLiquidChromatography;GF=GravityFlow;HPLC=HighPerformanceLiquidChromatography;ICC=Immunocytochemistry;IF=Immunofluorescence;IHC=Immunohistochemistry;IP=Immunoprecipitation;NAC=Non-adherentCellAssays;NB=NeutralizationofBioactivity;SE=SandwichELISA;TPE=TargetedProteinExpression;WB=Westernblotting;;AC=AdherentCellAssays;FM=FluorescentMicsroscopy;;;BSC-CM5=BiacoreSensorChipCM5;BSM=BiosactiveSmallMoleculeorPeptide;CDM=CellDifferentiationMedia;;;;;;HealthandFitness;;;DNAExtraction/Purification;;InvivoLikeAssaysSpeciesReactivity:H;M;Pr;R
B=Bovine;Ca=Cat;Ch=Chicken;D=Dog;EQ=Equine;GP=GuineaPig;H=Human;M=Mouse;P=Porcine;Pr=Primate;R=Rat;Rb=Rabbit;Y=Yeast;Xe=Xenopus;Ze=Zebrafish;;;;NA-NotApplicable;STP=Step-TactinProteins;AllFormat:AffinityPurified-liquid
Immunogen:NHQLENLEAETAPLP
Threetypesofopioidreceptorshavebeencloned--mu,delta,andkappa.OpioidreceptorsareseventransmembraneG-proteincoupledreceptors.TheyshareahighdegreeofhomologyandaremostdivergentattheN-andC-termini.ActivationofmuopioidreceptorsleadstoadecreaseinneuronalexcitABIlity.
Image:MuOpioidReceptorstainingofRatDorsalHorn.CourtesyofDr.LouisGendron,UniversityofSherbrooke.
Protocol:
Ratswere deeplyanesthetized withisoflurane andperfusedthroughtheaorticarchwith 100mlofheparin(75U/mlofheparinin0.9%saline)followedby100mlofamixtureof3.75%acroleinand2%PFAin0.1MPB,pH7.4,andthenby300mlof2%PFAinthesamebufferat45ml/min.mLumbar spinalcordwas removedandpostfixedin2%PFAin0.1MPBfor1hat4°C.Sections(50µmthick)werecutusingaVibratomeandprocessedforMORlabeling.
Sectionswereincubatedin1%sodiumborohydridefor30minandextensivelyrinsedin0.1MPB.Theywerethencryoprotectedfor30mininasolutionconsistingof25%sucroseand3%glycerolin0.05MPBandsnapfrozenwithisopentane(–50°C)followedbyliquidnitrogen.
Afterbeingrapidlythawedin0.1MPB,sectionswererinsedwithTBS 0.1M andpreincubatedfor1hatroomtemperaturein3%NGSdilutedinTBS.Theywerethenincubatedfor36hat4°CinMORantiserum diluted 1/500inTBScontaining0.5%NGS.SectionswerethenrinsedtwicewithTBSandincubatedfor1hatroomtemperaturewithbiotinylatedanti-guineapigantibody(1/400;VectorLaboratories).
Followingthree10minwashesinTBS,sectionswereincubated30minwith VectastainEliteABC(VectorLaboratories).SectionswererinsedthreetimeswithTBSandperoxidasecomplexrevealedfor8minuteswithDABsubstrate(2.2mg/10ml+0.01%H2O2).
Attheendofthisincubation,sectionswerewashedtwicewithTBS, mountedonmicroscopeslides,anddehydratedwithethanol.
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