Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification.

ABSTRACT: Absolute quantification of target proteins within complex biological samples is critical to a wide range of research and clinical applications. This protocol provides step-by-step instructions for the development and application of quantitative assays using selected reaction monitoring (SRM) mass spectrometry (MS). First, likely quantotypic target peptides are identified based on numerous criteria. This includes identifying proteotypic peptides, avoiding sites of posttranslational modification, and analyzing the uniqueness of the target peptide to the target protein. Next, crude external peptide standards are synthesized and used to develop SRM assays, and the resulting assays are used to perform qualitative analyses of the biological samples. Finally, purified, quantified, heavy isotope l

SUBMITTER: Manes NP 

PROVIDER: S-EPMC4692538 | biostudies-literature | 2015 Aug

REPOSITORIES: biostudies-literature