Abstract
Shotgun proteomics has a key role in quantitative estimation of proteins from biological systems under different conditions, which is crucial in the understanding of their functional roles. Isobaric tagging for relative and absolute quantitation (iTRAQ) mass spectrometry is based on pre-labeling of peptides with mass tags which allows the multiplex analysis of up to eight proteomes simultaneously. We describe here a detailed protocol for sample preparation and iTRAQ 4-plex labeling for relative quantification of multiple samples from human and plant tissues. We also present two strategies for peptide fractionation after the iTRAQ labeling protocol.
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Acknowledgments
This work was funded by the CNPq (Grant number: 477325/2013-0) and FAPERJ (Grant number: E-26/202.801/2015).
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Núñez, E.V., Domont, G.B., Nogueira, F.C.S. (2017). iTRAQ-Based Shotgun Proteomics Approach for Relative Protein Quantification. In: Guest, P.C. (eds) Multiplex Biomarker Techniques. Methods in Molecular Biology, vol 1546. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-6730-8_23
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DOI: https://doi.org/10.1007/978-1-4939-6730-8_23
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