Immunoproteomics using polyclonal antibodies and stable isotope-labeled affinity-purified recombinant proteins.
The combination of immuno-based methods of detection and mass spectrometry has great potential in the field of quantitative proteomics. Here, we describe a new method (immuno-SILAC) for the absolute quantification of proteins in complex samples by polyclonal antibodies and stable isotope-labeled recombinant protein fragments to allow the affinity enrichment prior to mass spectrometry analysis and accurate quantification. We took advantage of the resources available antibodies to the public from the Human Protein Atlas project covers more than 80% of all human protein-coding genes. Epitope mapping revealed that the majority of polyclonal antibodies recognized multiple linear epitopes, and based on these results, semi-automated method was developed for the enrichment of peptides using polyclonal antibodies immobilized on magnetic beads to protein A-coated . Swine Recombinant Proteins A protocol based on the arrest of more than 40 simultaneous multiplex protein targets show tha...