Description:C3b/iC3b/C3c,Mouse,mAb2/11,immunoassayantibodyThemonocolonalantibody2/11recognizesactivatedfragmentsofthemousecomplementproteinC3.Thecomplementsystemisanimportantpartofthehumoralresponseininnateimmunity,consistingofthreedifferentpathways.Thethirdcomplementcomponent,C3,iscentraltotheclassical,alternativeandlectinpathwaysofcomplementactivation.Activationproductsofthecomplementcascadecontainneo-epitopesthatarenotpresentintheindividualnativecomponents.ThesynthesisofC3istissue-specificandismodulatedinresponsetoavarietyofstimulatoryagents.C3isthemostabundantproteinofthecomplementsystemwithserumproteinlevelsofabout1.3mg/ml.AninheriteddeficiencyofC3predisposesthepersontofrequentassaultsofbacterialinfections.Inulcerativecolitis,andidiopathicchronicinflammatoryboweldisease,thedepositionofC3inthediseasedmucosahasbeenreported.ProteolysisbycertainenzymesresultsinthecleavageofC3intoC3aandC3b.C3bbecomesattachedtoimmunecomplexesandisfurthercleavedintoiC3b,C3c,C3dgandC3f.Themonoclonalantibody2/11isspecificforcleavedC3fragmentsC3b,iC3b,andC3c,andforactivatedC3.Therefore,positivereactivityintissuesisassociatedwithactivationofhecomplementcascadeandC3cleavage.IncaseofanacuteinflammatoryreactionlotsofC3areprocessedintotheproductsrecognizedby2/11andisassuchusefulasMarkerforinflammatoryreaction.Inchronicinflammatoryconditionsminimalreactivitywith2/11mayobserved.InsuchcasesprimarilytheC3dgproductresidesattheplaceofinflammation(C3cbeingcleared)whichisnotrecognizedbyantibody2/11.Thechronicprocessing/activationofC3istakingplaceatalowerlevel,whichwouldreducedetectionoftheC3fragmentsC3b,iC3b,andC3c.NexttodetectionofC3fragmentsC3b,iC3b,andC3c,themonoclonalantibody2/11inhibitsthehemolyticactivityofmousecomplementinadose-dependentmanner.Specifications:CatalognumberHM1065-IAProducttypeMonoclonalantibodiesQuantity0.5mgFormulation0.5mgof0.2μmfilteredproteinGpurifiedantibodysolutioninPBSwithaconcentrationofatleast0.5mg/ml(exactconcentrationisindicatedonthelabel).ApplicationFlowcytometry,Frozensections,Functionalstudies,Immunoassays,Immunofluorescence,WesternblotApplicationNotesF:Fixationofsectionsinacetonefor5‘,pretreatmentwith3%hydrogenperoxideinmethanolatRTtoquenchendogenousperoxidases.Asnegativecontrol,sectionsfromC3-deficientmicewereused.(Ref.1)FC:Mousesplenocyteswereincubatedwithautologous,freshlydrawnmouseserumtofixtheC3fragmentsbeforestaining.AsnegativecontrolsplenocytesofC3deficientmicewasused.Antibody2/11stainstheneoantigenicsiteontheC3activationfragments.(Ref.1)FS:Antibody2/11inhibitshemolysisinadose-dependentmanner.(Ref.1)W:Anon-reducedsampletreatmentandSDS-PAGEwasused.Thebandsizeis150kDa(Ref.1).IA:Supernatantofantibody2/11wasincubatedonanti-ratIgGFccoatedplates.Afterincubationwithsample,HRP-conjugatedpolyclonalgoatanti-mouseC3wasusedfordetection.(Ref.1)UseDilutionstobeuseddependondetectionsystemapplied.Itisrecommendedthatuserstestthereagentanddeterminetheirownoptimaldilutions.Thetypicalstartingworkingdilutionis1:50.ImmunogenMouseC3IsotypeRatIgG1SpeciesMousePositiveControlMousesplenocytespreincubatedwithserumfromthesameanimal.NegativeControlC3-deficientmousecells.References1.Mastellos,Detal;NovelmonoclonalantibodiesagainstmouseC3interferingwithcomplementactivation:descriptionoffinespecificityandapplicationstovariousimmunoassays.MolImmunol2004,40:12132.Markiewski,Metal;C3aandC3bactivationproductsofthethirdcomponentofcomplement(C3)arecriticalfornormalliverrecoveryaftertoxicinjury.JImmunol2004,173:7473.Markiewski,Metal;Modulationoftheantitumorimmuneresponsebycomplement.NatImmunol2008,9:1225StorageandstABIlityProductshouldbestoredat4 °C.Underrecommendedstorageconditions,productisstableforoneyear.PrecautionsForresearchuseonly.Notforuseinoronhumansoranimalsorfordiagnostics.Itistheresponsibilityoftheusertocomplywithalllocal/stateandFederalrulesintheuseofthisproduct.HycultBiotechisnotresponsIBLeforanypatentinfringementsthatmightresultwiththeuseoforderivationofthisproduct.DiseaseInfectiousdiseases,Nephrology