Abstract
High-field asymmetric waveform ion mobility spectrometry (FAIMS) is a gas-phase separation technique which, when coupled with liquid chromatography tandem mass spectrometry, offers benefits for analysis of complex proteomics samples such as those encountered in phosphoproteomics experiments. Results from LC-FAIMS-MS/MS are typically complementary, in terms of proteome coverage and isomer identification, to those obtained by use of solution-phase separation methods, such as prefractionation with strong cation-exchange chromatography. Here, we describe the protocol for large-scale phosphorylation analysis by LC-FAIMS-MS/MS.
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Acknowledgments
The Advion Triversa Nanomate, Dionex LC and Thermo Fisher Velos Orbitrap mass spectrometer used in this research were funded through the Birmingham Science City Translational Medicine: Experimental Medicine Network of Excellence Project, with support from Advantage West Midlands (AWM). The Chinese Scholarship Council is gratefully acknowledged for funding. HJC is an EPSRC Established Career Fellow (EP/L023490/1).
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Zhao, H., Creese, A.J., Cooper, H.J. (2016). Online LC-FAIMS-MS/MS for the Analysis of Phosphorylation in Proteins. In: von Stechow, L. (eds) Phospho-Proteomics. Methods in Molecular Biology, vol 1355. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-3049-4_16
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DOI: https://doi.org/10.1007/978-1-4939-3049-4_16
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4939-3048-7
Online ISBN: 978-1-4939-3049-4
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