MakingAntibody

·ProductionofPolyclonalAntibodyinRabbit(WalterSteffen)Providesdetailedprotocolforimmunizatioin,bleedingprocedure,obtainingserum,precipitationofIgG.

·GST-fusionproteininjectionintorabbits(IVANJ.DELGADOORLIC)

·HowDoImakeanAntibodyagainstaPeptides(Pierce)

·Afastandinexpensivemethodforraisinghigh-specificitypolyclonalantibodiesinmice(TechnicalTipsOnline)

·GuidetoAntipeptideAntibody(Perkin-Elmer)GeneralInformationaboutthedesignandsynthesisofpeptidesfortheproductionofanti-peptideantibodies,aswellasanoverviewofthestepsinvolvedintheproductionofantibodies­immunization,collectionofantisera,analysisofantisera,andantibodypurification.Protocolsforanumberoftheseproceduresareprovided.

oDesigningAntigens

oSynthesizingPeptides

oConjugationofPeptidestoProteinCarriers

oImmunizations

oAnalysisofAntisera:Immunoassays

oPurifyingAntibodies

• PreparationofPolyclonalAntibodies(PMCI)Immunizationandantiserapreparation
• Makingantibodiestosyntheticpeptides(DanielT.O"Connor)
• PreparationofPeptide-KLHConjugatesforImmunization(MitshisonLab)
• TestingRabbitBleedsbyELISA(MitshisonLab)
• CouplingPeptidestoResinforAffinityPurification(MitshisonLab)
• PurificationofAntiPeptideAntibody(MitshisonLab)

Others

·Tipsandhintsforthestorageofantibodies(SynapticSystems)

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