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Using 15N-Metabolic Labeling for Quantitative Proteomic Analyses.

MPS-Authors
http://pubman.mpdl.mpg.de/cone/persons/resource/persons80429

Maccarrone,  Giuseppina
Dept. Translational Research in Psychiatry, Max Planck Institute of Psychiatry, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons129892

Chen,  Alon
Dept. Stress Neurobiology and Neurogenetics, Max Planck Institute of Psychiatry, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons80323

Filiou,  Michaela D.
Dept. Stress Neurobiology and Neurogenetics, Max Planck Institute of Psychiatry, Max Planck Society;

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Citation

Maccarrone, G., Chen, A., & Filiou, M. D. (2017). Using 15N-Metabolic Labeling for Quantitative Proteomic Analyses. Methods in molecular biology (Clifton, N.J.), 1546, 235-243.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002D-78AB-6
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.