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Free keywords:
15N-Metabolic labeling; Quantitative Proteomics; Mass spectrometry; Peptide quantification
Abstract:
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 (15)N 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.
