Immunoproteomics using polyclonal antibodies and stable isotope-labeled affinity-purified recombinant proteins.

ABSTRACT: The combination of immuno-based methods and mass spectrometry detection 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 based on polyclonal antibodies and stable isotope-labeled recombinant protein fragments to allow affinity enrichment prior to mass spectrometry analysis and accurate quantification. We took advantage of the antibody resources publicly available from the Human Protein Atlas project covering more than 80% of all human protein-coding genes. Epitope mapping revealed that a majority of the polyclonal antibodies recognized multiple linear epitopes, and based on these results, a semi-automated method was developed for peptide enrichment using polyclonal antib

SUBMITTER: Edfors F 

PROVIDER: S-EPMC4047479 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature