Description | MonoclonalantibodyMC192againsttheratlowaffinitynervegrowthfactorreceptor(p75NTR)isderivedfromthefusionofSp2/0-Ag14myelomacellswithmouseimmunesplenocytes.MC192monoclonalantibodywasoriginallygeneratedbyChandlersetal.p75NTRwasoriginallydiscoveredasalowaffinitynervegrowthfactorreceptor.Lateritwasfoundthatitwasthereceptorforallneurotrophins.Itmediatessignalsofneurotrophinsforneuronalsurvival,apoptosis,neuriteoutgrowthandsynapticplasticity.Recently,ithasbeenrevealedthatp75NTRnotonlyactsasthereceptorforneurotrophinsbutalsothereceptorformanyotherpathologicalligandssuchasprions,rABIesvirusandamyloidbeta.p75NTRalsoactsasaco-receptorforNOGOwhichmediatesinhibitorysignalsofmyelinassociatedprotein.p75NTRishighlyexpressedinanumberofnon-neuronalandneuronalcellsincludingmotorneuronsduringdevelopmentandalsoindamagedneurons.MC192recognizestheextracellulardomainoftheneurotrophinreceptorp75NTRinrat.MC192antibodymaybeusedforimmunocytochemicallocalisationofratcellsexpressingp75NTR,ELISAandwesternblot.ThisantibodyhasalsobeenusedfortheconstructionoftheMC192-saporinimmunotoxinforspecificeliminationofneuronalpopulationsinbasalforebraincholinergicneuronstogenerateananimalmodelforAlzheimer"sdisease.UsingFlowCytometry,thisantibodyhasfrequentlybeenemployedforpanningtoisolatep75NTR-expressingratcells.MC192hasapotentialuseastheligandforgenedeliveryintop75NTR-expressingratcellsviaareceptor-mediatedmechanism.FUNCTION:LowaffinityreceptorwhichcanbindtoNGF,BDNF,NT-3,andNT-4.Canmediatecellsurvivalaswellascelldeathofneuralcells.SUBUNIT:Homodimer;disulfide-linked.Interactswithp75NTR-associatedcelldeathexecutor.InteractswithNGFRAP1/BEX3.InteractswithTRAF2,TRAF4,TRAF6,PTPN13andRANBP9.InteractsthroughTRAF6withSQSTM1whichbridgesNGFRtoNTRK1(Bysimilarity).InteractswithBEX1.SUBCELLULARLOCATION:Membrane;single-passtypeImembraneprotein.DOMAIN:DeathdomainisresponsIBLeforinteractionwithRANBP9.PTM:N-andO-glycosylated.PTM:Phosphorylatedonserineresidues.SIMILARITY:Contains1deathdomain.SIMILARITY:Contains4TNFR-Cysrepeats. |
BatchNo. | Seeproductlabel |
Unitsize | 100µg |
Antigen | NGFreceptor |
AntigenLocation | ECDofratNGFR |
Isotype | IgG1 |
Clone | MC192 |
OtherNames | Low-affinitynervegrowthfactorreceptor;NGFreceptor;Gp80-LNGFR;p75ICD;Lowaffinityneurotrophinreceptorp75NTR |
Accession | TNR16_RAT |
Producedin | Mouse |
Purity | ProteinGpurifiedimmunoglobulin |
Applications | IH(likelyfixed),ELISA,WB,FlowCytometry(2µgper10^6cells)IP(non-reducingconditionsonly!;donotusereducingagentssuchasDTTorbeta-mercaptoethanol),TraditionalformalinfixedparaffinembeddedimmunohistochemistryisNOTrecommendedwithMC192.Motorneuronisolation,Gene/ToxinDeliverytoratsensory/motorneurons.Aworkingsolutionof1-2µg/mlwasdeterminedbyimmunohistochemicalstainingon4%paraformaldehydefixed,oralcoholfixedratspinalcordandbrain.Fornon-denaturedWB,1-5ug/mLwasfoundtobesuitablewithsuitablecontrols(PC12lysate).ELISA:detectiononly,1-5ug/mLhasbeensuggestedinliterature.Immunoprecipitation:5μg/mL,>0.5%tritonX-100buffer/500ug/lysate;PC12positivecontrolstrongsuggested.MC192isnotsuitableasablockingagent,althoughithasbeenincorrectlyusedforthispurposeinmanypublishedworks.TheantibodywasgeneratedspecificallybyscreeningformonoclonalsthathadtheabilitytoENHANCEthebindingofNGF,thenaturalligandforp75.Therefore,thisantibodyisparticularlyunusual.Thefulldetailscanbefoundintheoriginalpaper,whichislistedonourdatasheet(seeChandleretal,1984).Biosensisrecommendsoptimaldilutions/concentrationsshouldbedeterminedbytheenduser. |
Specificity | MC192isspecificonlyforRATNGFR,noreactivitytoHumanorMouseNGFRhasbeenreported |
Cross-reactivity | Thismonoclonalantibodyhasbeentestedforimmunohistochemicallocalisationofp75NTR-expressingratcellsinthespinalcordandbrain.Thismonoclonalantibodydoesnotcrossreactwithp75NTR-expressingcellsinotherspecies. |
Form | Lyophilised |
Reconstitution | Reconstitutein100µlofsterilewater.Centrifugetoremoveanyinsolublematerial. |
Storage | TheMC192issuppliedinlyophilisedformfromProteinG-purifiedhybridomacellculturesupernatants.Thelyophilisedantibodyisstablewhenstoredat2-8°Cor-20°C.Afterreconstitutionundilutedaliquotsshouldbekeptat-20°Cforuptosixmonths.ForadditionalstabilityGlycerol(1:1)maybeaddedafterreconstitution.Repetitivefreeze/thawcycleshouldbeavoided. |
SpecificReferences | RiffaultB,KourdougliN,DumonC,FerrandN,BuhlerE,SchallerF,ChambonC,RiveraC,GaiarsaJL,PorcherC(2016)Pro-Brain-DerivedNeurotrophicFactor(proBDNF)-Mediatedp75NTRActivationPromotesDepolarizingActionsofGABAandIncreasesSusceptibilitytoEpilepticSeizures.Cereb.Cortex[Epubaheadofprint].Application:WesternBlot;Species:Rat BrandliA,JohnstoneDM,StoneJ(2016)RemoteIschemicPreconditioningProtectsRetinalPhotoreceptors:EvidenceFromaRatModelofLight-InducedPhotoreceptorDegeneration.InvestOphthalmolVisSci.57(13):5302-13Application:WesternBlot,IHC;Species:Rat RiffaultB,MedinaI,DumonC,ThalmanC,FerrandN,FriedelP,GaiarsaJL,PorcherC.(2014)"Pro-Brain-DerivedNeurotrophicFactorInhibitsGABAergicNeurotransmissionbyActivatingEndocytosisandRepressionofGABAAReceptors."J.Neurosci.34(40):13516-34Application:WesternBlot,Neuronalcellsandhippocampi;Species:Rat KalincikTetal(2011)SelectedchangesinspinalcordmorphologyafterT4transectionandolfactoryensheathingcelltransplantation.AutonNeurosci.158(1-2):31-8Application:IF;Species:Rat WuAetal(2011)Delayedolfactoryensheathingcelltransplantsreducenociceptionafterdorsalrootinjury.ExpNeurol.229(1):143-57Application:IF;Species:Rat DaviesAetal(2010)Thealpha2deltasubunitsofvoltage-gatedcalciumchannelsformGPI-anchoredproteins,aposttranslationalmodificationessentialforfunctionProcNatlAcadSciUSA.Jan26;107(4):1654-9 KalincikTetal(2010)Olfactoryensheathingcellsreducedurationofautonomicdysreflexiainratswithhighspinalcordinjury.AutonNeurosci.154(1-2):20-9Application:IHC;Species:Rat Wilson-GerwingT.D.etal(2009)JCompNeurol.2009Sep1;516(1):49-58 FeronFetal(2008)Neurotrophinexpressionintheadultolfactoryepithelium.BrainRes.1196:13-21Application:IHC;Species:Rat BiancoJIetal(2004)Neurotrophin3promotespurificationandproliferationofolfactoryensheathingcellsfromhumannose.Glia.45(2):111-23Application:IHC,IF;Species:Rat EylesDetal(2003)Neuroscience.2003;118(3):641-53.Application:IHC;Species:Rat LuJetal(2001)Transplantationofnasalolfactorytissuepromotespartialrecoveryinparaplegicadultrats.BrainRes.889(1-2):344-57Application:IF;Species:Rat |
GeneralReferences | VilarM.etal(2009)Activationofthep75neurotrophinreceptorthroughconformationalrearrangementofdisulphide-linkedreceptordimersNeuron.2009Apr16;62(1):72-83 Luther,J.A.andBirren,S.J.(2009)J.Neurosci29,5411-24 Lagares,A.etal.(2007)J.Neurosci27,7939-53 Kruger,G.M.etal.(2002)Neuron35,657-69 Tuffereau,C.etal(1998)EmboJ17,7250-9 Zhou,X.F.,andRush,R.A.(1996)JCompNeurol372,37-48 Huber,J.,Dittrich,F.,andPhelan,P.(1993)EurJBiochem218,1031-1039 RabizadehS.etal(1993)Science.Jul16;261(5119):345-8. Stemple,D.L.&Anderson,D.J.(1992)Cell71,973-85 Urschel,B.A.&Hulsebosch,C.E.(1992)BrainResDevBrainRes69,261-70 BirrenSJetal(1992)Science.Jul17;257(5068):395-7. Yan,Q.,andJohnson,E.M.,Jr.(1988)JNeurosci.1988Sep;8(9):3481-98. Johnson,D.etal.(1986)Cell47,545-54 Chandler,C.E.etal(1984)(1984)JBiolChem259,6882-6889 |
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