Using 15 N-Metabolic Labeling for Quantitative Proteomic Analyses

: Quantitative proteomics has benefited from the application of stable isotope labeling-based approaches. Using stable isotopically labeled material as an internal standard in proteomic comparisons allows an unbiased and accurate quantification of protein expression level changes. Here, we describe the use of in vivo 15N metabolic labeling to generate labeled protein standards from mice. We then present a protocol including sample preparation, mass spectrometry, and data analysis workflows using these standards to compare unlabeled proteomes. We focus on mouse brain tissue and plasma samples, although this conceptual framework can be applied to most organisms.