Description:C5b-9,Rat,mAb2A1,immunoassayantibodyThemonoclonalantibody2A1recognizesratC5b-9.TheantibodywasshowntocompetewithantibodiestohumanC9foritsbindingsiteontheC5b-9complex,indicatingthatthereactiveepitopeislocatedontheC9molecule.C5b-9membraneattackcomplexesareassembledfromfiveprecursormoleculesintheserum.ProteolyticcleavageofC5byC5convertasegeneratesC5bwhichinitiatesassemblyoftheC5b-9complex.ThelaststepofC5b-9complexformationinvolvespolymerizationofC9whichaccompaniesinsertionofthecomplexintothecellmembrane.DuringformationofC5b-8andC9polymerization,neoantigensaregeneratedwhichareuniquetotheC5b-9complexandarenotpresentonanyoftheindividualnativecomplexcomponents.ThecomplementregulatoryproteinsCD59andcomplementS-proteincanbothpreventC5b-9insertionintothecellmembrane.TheformedSC5b-9complexisunabletoattachtocellsandiscytolyticallyinactive.C5b-9isinvolvedintheprogressionofchronicproteinuricrenaldiseasebymediatingcontinuoustubulointerstitialdamage.EarlytubulointerstitialinjuryintheremnantkidneycanbeimprovedwhenC5b-9complexformingisabrogated.Themonoclonalantibody2A1wasraisedagainstaratC5b-9neoantigen.Monoclonalantibody2A1canbeusedasacoatingantibodytodetectC5b-9inplasmaandurinesamples.Specifications:CatalognumberHM3033-IAProducttypeMonoclonalantibodiesQuantity0.5mgFormulation0.5mgof0.2μmfilteredproteinGpurifiedantibodysolutioninPBS(exactconcentrationisindicatedonthelabel).ApplicationFlowcytometry,Frozensections,Immunoassays,Immunofluorescence,Paraffinsections,WesternblotApplicationNotesIA:plateswerecoatedwith30μg/mlin50mMcarbonatebuffer,pH10.6for16hat4°C.(Ref.1)W:Anon-reducedsampletreatmentandSDS-Pagewasused.Thebandsizeis>200kDa(Ref.1).IHC-P:TissuesectionsfixedinformalinwerepretreatedwithproteasetypeXXIVfor10minutesat37⁰Cbeforeincubation(Ref.6).IHC-F:Tissuesectionswerefixedinacetonefor10minutesatroomtemperaturebeforeincubation(Ref.3).UseDilutionstobeuseddependondetectionsystemapplied.Itisrecommendedthatuserstestthereagentanddeterminetheirownoptimaldilutions.Thetypicalstartingworkingdilutionis1:50.ImmunogenRatC5b-9IsotypeMouseIgG1SpeciesRatAliasMembraneattackcomplex,MAC,TCC,TerminalComplementComplexPositiveControlGlomeruliofratstreatedwithanti-Thy-1.1antibodiesReferences1.Schulze,Metal.IncreasedurinaryexcretionofC5b-9distinguishespassiveHeymannnephritisintherat.KidneyInt.1989,35:602.Brandt,Jetal.Roleofthecomplementmembraneattackcomplex(C5b-9)inmediatingexperimentalmesangioproliferativeglomerulonephritis.KidneyInt.1996,49:3353.Sato,Tetal.Theterminalsequenceofcomplementplaysanessentialroleinantibody-mediatedrenalcellapoptosis.JAmSocNephrol1999,10:12424.Duijvestijn,A.Complementactivationbyanti-endothelialcellantibodiesinMHC-mismatchedandMHC-matchedheartallograftrejection:anti-MHC,butnotantinon-MHCalloantibodiesareeffectiveincomplementactivation.Transpl.Int.2000,13:3635.Yamada,Ketal.Clusterinisup-regulatedinglomerularmesangialcellsincomplement-mediatedinjury.KidneyIntern2001.59:1376.NangakuMetal.C6mediateschronicprogressionoftubulointerstitialdamageinratswithremnantkidneysJ.Am.Soc.Nephrol.2002,13:9287.RanganGKetal.C5b-9regulatesperitubularmyofibroblastaccumulationinexperimentalfocalsegmentalglomerulosclerosis.KidneyInt2004,66:18388.Ostendorf,Tetal.AntagonismofPDGF-DbyhumanantibodyCR002preventsrenalscarringinexperimentalglomerulonephritis.JAmSocNephrol2006,17:10549.Pol,P;Pathogenicroleofcomplementinrenalischemia/reperfusioninjury.Thesis201310.Kotimaa,Jetal;FunctionalassessmentofratcomplementpathwayactivitiesandquantificationofsolubleC5b-9inanexperimentalmodelofrenalischemia/reperfusioninjury.JImmMeth2014,412:14StorageandstABIlityProductshouldbestoredat4 °C.Underrecommendedstorageconditions,productisstableforoneyear.PrecautionsForresearchuseonly.Notforuseinoronhumansoranimalsorfordiagnostics.Itistheresponsibilityoftheusertocomplywithalllocal/stateandfederalrulesintheuseofthisproduct.HycultBiotechisnotresponsIBLeforanypatentinfringementsthatmightresultfromtheuseorderivationofthisproduct.DiseaseInfectiousdiseases,Nephrology