Background:

Inimmunoelectrophoresisagainstfreshguineapigserum,asingleprecipitinlineisobtainedinthebeta-1regionrepresentingnativeC3.AgainstserumcontainingpartlyactivatedC3,aprecipitinlineisobtainedwhichextendsfromthebeta-1intothealpha-2region,demonstratingagrADIent.InoldserumcontainingtotallyactivatedC3asingleprecipitinlineinthealpha-2regionisobtained.AntiseratoC3ccabalsoreactwiththefragmentsC3b,C3biandsmallerfragments,sincetheyallcarryantigenicdeterminantsoftheC3cdomain.Theproductdoesnotreactwithanyotherproteinscomponentofguineapigserumorplasma.ThebiotinimmunoconjugatetoguineapigC3cisusedtodeterminethepresenceandpatternofC3intissuelesionsusingimmunohistochemicalstainingtechniques.Locallydepositedimmunecomplexesintissueusuallycontaincomplement,pointingtoactivationoftheclassicalpathway.Complementactivationinvivoimpliesactivediseaseandmaycontributetotheelicitationofthepathogenesisandheextentoftissuedestruction.InELISAandWesternblottingtoidentifyguineapigC3cinserumorotherbodyfluids.Asasecondstepanavidinorstreptavidinconjugateoftheuser’schoicehastobeused.Thisimmunoconjugateisnotpre-diluted.Theoptimumworkingdilutionofeachconjugateshouldbeestablishedbytitrationbeforebeingused.Excesslabelledantibodymustbeavoidedbecauseitmaycausehighunspecificbackgroundstainingandinterferewiththespecificsignal.

Source:

C3isthemostabundantcomplementproteininguineapigserum.ItsBIOLOGicalfunctionstronglyresemblesthatofC3inmananotherlaboratoryanimalspecies.Ithasacentralroleintheactivationsystembeingcommontobothpathways.ActivationofC3isachievedbyveryspecificlimitedproteolysisresultinginthereleaseofanumberofdegradationfragments.TheanaphylotoxinC3apromotessmoothmusclecontractionandincreasesvascularpermeABIlity:thelargeC3bfragmentisinvolvedinbindingtothecomplementactivatorandcanbeinteractwithspecificreceptorstoallowefficientclearanceoftheactivatingcellorparticle;degradationfragmentsofC3b(C3bi,C3c,C3dgC3d)areimportantinreceptorbindingandclearancemechanisms,invirusneutralizationandpossIBLyintheimmuneresponse.TheantiserumisraisedagainstC3c,whichisthemajorfragmentresultingfromC3cleavagebyC3convertaseandfactorI.Itiscomposedofanintactbetachainboundtotwofragmentsofthealphachain.ConsequentlytheantiserumreactswithbothnativeandactivatedC3.ItmayalsoreactwiththefragmentsC3b,C3biandC3dg,sincetheyallcarryantigenicepitopesoftheC3cdomain.C3cisisolatedandpurifiedfrompoolednormalguineapigserum.Freund’scompleteadjuvantisusedinthefirststepoftheimmunizationprocedure.

Product:

Biotin-coupledpurifiedhyperimmuneIgGlyophilizedfromasolutioninphosphatebufferedsaline(PBS,pH7.2)Nopreservativeadded,asitmayinterferewiththeantibodyactivity.Itisreconstitutedbyadding1mlsteriledistilledwater,spundowntoremoveinsolubleparticles,dividedintosmallaliquots,frozenandstoredatorbelow-20°C.

Applications:

ELISA,Immunocytochemistry,Immunohistochemistry(paraffin),Dotblot,Immunoblotting.

CrossReactivity:

TheantiSerumdoesnotcross-reactwithanyothercomponentofGuineaPigplasma.Inter-speciescross-reactivityisanormalfeatureofantibodiestoplasmaproteinssincetheyfrequentlyshareantigenicdeterminants.Cross-reactivityofthisantiSerumhasnotbeentestedindetail.

Specificity:

Biotin-conjugatedIgGfractionofpolyclonalGoatantiSerumtoC3cfragmentofGuineaPigcomplementfactorC3

Storage:

Thelyophilizedconjugateisshippedatambienttemperatureandmaybestoredat+4°C;prolongedstorageatorbelow-20°C.Priortouse,analiquotisthawedslowlyatambienttemperature,spundownagainandusedtoprepareworkingdilutionsbyaddingsterilephosphatebufferedsaline(PBS,pH7.2).Repeatedthawingandfreezingshouldbeavoided.Workingdilutionsshouldbestoredat+4°C,notrefrozen,andpreferablyusedthesameday.Ifaslightprecipitationoccursuponstorage,thisshouldberemovedbycentrifugation.Itwillnotaffecttheperformanceoftheimmunoconjugate.

Caution:

ThisproductisintendedFORRESEARCHUSEONLY,andFORTESTSINVITRO,notforuseindiagnosticortherapeuticproceduresinvolvinghumansoranimals.Thisdatasheetisasaccurateasreasonablyachievable,butNordic-Mubioacceptsnoliabilityforanyinaccuraciesoromissionsinthisinformation.

Noridic-Mubio是荷兰的一家著名的免疫学试剂生产厂商，因收购An Der Grub Bio Research GmbH(ADG)，增加了自己的产品线：全球最好的固定破膜剂FIX&PERM 。Nordic-Mubio严格遵照ISO9001标准生产高质量的抗体，主要为针对细胞粘附蛋白、核蛋白和骨架蛋白的抗体。同时Nordic-Mubio也是斑马鱼抗体开发的引领者之一，拥有50多种可用于斑马鱼免疫荧光检测的抗体。