Description:C5,Mouse,mAbBB5.1,immunoassayantibodyThemonoclonalantibodyBB5.1bindsthefifthcomponentofmousecomplement(C5).Thecomplementsystemisagroupofplasmaandcellmembraneproteinsthatplayakeyroleintheimmunesystem.Allthreepathways(classical,alternativeandlectin)leadtothecleavageofC3andeventuallytheformationofthecytolyticmembraneattackcomplexC5b-9.IftheactivationcascadeisallowedtoproceedbeyondthecleavingofC3intoC3aandC3b,anadditionalC3bmoleculebindstotheC3convertases.ThisgeneratestheC5convertase(C3bBbC3bforthealternativepathwayandC4bC2bC3bfortheclassicalandlectinpathways).C5convertasecleavesC5,releasesthepotentanaphylacticpeptideC5aandgeneratesC5b.C5bcaninitiatetheterminalpathway,whichrecruitsthecomponentsC6,C7,C8andC9tothesurfaceofthetargetandinsertstheC9complexasapore(termedtheterminalcomplementcomplex)intothemembrane.C5aisthemostpotentanaphylatoxinandapowerfulchemotaxinforneutrophilsandmonocytes,withtheABIlitytopromotemargination,extravasation,andactivationofthesecells.Thus,blockingC5mayberequiredforoptimalinhibitionoftheinflammatoryresponse.Atthesametime,inhibitionofthecomplementcascadeatC5doesnotimpairthegenerationofC3bthroughtheclassicalandalternativepathways,preservingC3b-mediatedopsonizationofpathogenicmicroorganismsaswellasopsonizationandsolubilizationofimmunecomplexes.C5issynthesisedintheliverasasinglepolypeptidechainandispresentinseruminaconcentrationof50-80 µm/ml.Beforesecretionthemoleculeisglycosylatedandsecretedintoplasmaasa190kDaglycoproteinconsistingofadisulphidelinkedalpha-chain(111kDa)andbeta-chain(75kDa).MonoclonalantibodyBB5.1hasbeenshowntoprecipitatethetwochainsofC5fromnormalmouseserumandinhibitedC5-dependenthemolysisinafunctionalcomplementtest.FurThermore,BB5.1admiNISTrationcompletelyinhibitsterminalcomplementactivityinmurinemodelsforantibody-mediatedrejection(AMR)duringheartandkidneytransplantation.Inanothermousemodel,bothpre-treatmentaswellasinterventionwithmonoclonalantibodyBB5.1attenuatesdiseasedevelopmentduringanti-MPOIgG-inducedglomerulonephritis.BlockageofC5activationbyBB5.1protectsagainstrenalischemia-reperfusioninjurybyinhibitionoflateapoptosisandinflammation.InLupusdisease,combinationtherapyofanti-IL-10/anti-C5(BB5.1)canbothpreventandreducetheeffectofthehumoralimmuneresponse.Specifications:CatalognumberHM1073-IAProducttypeMonoclonalantibodiesQuantity0.5mgFormulation0.5mg(100μg/ml)0.2μmfilteredproteinGpurifiedantibodysolutioninPBSwithaconcentrationofatleast0.5mg/ml(exactconcentrationisindicatedonthelabel).ApplicationFrozensections,Functionalstudies,Immunoassays,Immunofluorescence,Immunoprecipitation,ParaffinsectionsApplicationNotesFS:exvivo:50 µgmAb/mlserum;invivo:BALB/cmice40mgmAb/kg/day,0-2daysfollowedbytwiceaweek.IA:Wellswerecoatedwith2-4 µgpurifiedBB5.1inPBSascoatantibodyforuseinELISA.IP:Mouseserumwaspre-adsorbedwithαmouse-Igaffinitygelandprot-A-sepharosefor30minatRT.ThenC5wasprecipitatedwithBB5.1(10 µgmAb/mlfollowedbyprot-Asepharosefor1h).Precipitatedsampleswereanalysedon7%SDS/PAGE.UseDilutionstobeuseddependondetectionsystemapplied.Itisrecommendedthatuserstestthereagentanddeterminetheirownoptimaldilutions.Thetypicalstartingworkingdilutionis1:50.ImmunogenPurifiedC5IsotypeMouseIgG1SpeciesMouseCrossreactivityRat-YesAliasComplementC5,HemolyticcomplementGenename:C5References1.Frei,Yetal;GenerationofamonoclonalantibodytomouseC5applicationinanELISAassayfordetectionofanti-C5antibodies.MolCellProbes1987,1:1412.Thurman,Jetal;C3aisrequiredfortheproductionofCXCchemokinesbytubularepithelialcellsafterrenalischemia/reperfusion.JImmunol2007,178:18193.Mihai,Setal;Thealternativepathwayofcomplementactivationiscriticalforblisterinductioninexperimentalepidermolysisbullosaacquisita.JImmunol2007,178:65144.Wang,Hetal;Inhibitionofterminalcomplementcomponentsinpresensitizedtransplantrecipientspreventsantibody-mediatedrejectionleADIngtolong-termgraftsurvivalandaccommodation.JImmunol2007,179:44515.Huugen,Detal;InhibitionofcomplementfactorC5protectsagainstantimyeloperoxidaseantibody-mediatedglomerulonephritisinmice.KidneyInt2007,71:6466.Nilsson,Ketal.Enhancedsusceptibilitytolow-dosecollagen-inducedarthritisinCR1/2-deficientfemalemice–possIBLeroleofestrogenonCR1expression.Faseb2009,23:24507.Copland,Detal.Systemicandlocalanti-C5therapyreducesthediseaseseverityinexperimentalautoimmuneuveoretinitis.Clinexpimmunol2010,159:3038.Pavlovski,Detal;GenerationofcomplementcomponentC5abyischemicneuronspromotesneuronalapoptosis,Faseb2012,26:3680StorageandstabilityProductshouldbestoredat4 °C.Underrecommendedstorageconditions,productisstableforoneyear.PrecautionsForresearchuseonly.Notforuseinoronhumansoranimalsorfordiagnostics.Itistheresponsibilityoftheusertocomplywithalllocal/stateandfederalrulesintheuseofthisproduct.HycultBiotechisnotresponsibleforanypatentinfringementsthatmightresultwiththeuseorderivationofthisproduct.DiseaseInfectiousdiseases,Nephrology