[20] Immunoprecipitation of human mitochondrial translation products with peptide-specific antibodies

Abstract

Publisher Summary This chapter describes the use of peptide-specific antibodies in immunoprecipitation experiments involving mtDNA-encoded polypeptides. It summarizes the steps carried out in producing the peptide-specific antibodies. Antibodies directed against a short synthetic peptide sequence predicted from a given gene sequence are able to recognize a full-length protein containing that short peptide sequence. This approach has been applied extensively for the identification of the proteins encoded in cloned genes. A successful application is in the identification of the mitochondrial translation products corresponding to the reading frames of human mitochondrial DNA (mtDNA). Peptide-specific antibodies specific for individual mtDNA-encoded polypeptides have also been used to immunoprecipitate entire respiratory complexes containing them and in the molecular analysis of defects associated with mitochondrial diseases.