Abstract
Pro-Q diamond phosphoprotein gel stain is a fluorescent stain to detect phosphorylated proteins in polyacrylamide gels with high sensitivity. Here, we describe an entire procedure for phosphoproteomics analysis of Arabidopsis seedlings by a combination of Pro-Q diamond stain and two-dimensional gel electrophoresis (2-DE). The workflow involves total protein preparation, protein separation by 2-DE, the second-dimensional gel staining, phosphoproteins detection, and peptides preparation for matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. Approximately 300 phosphoproteins can be detected using this method.
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Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (grant nos. 31670248 and 31970276) to D.R.
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Li, Y., Ren, D. (2021). Two-Dimensional Gel Electrophoresis and Pro-Q Diamond Phosphoprotein Stain-Based Plant Phosphoproteomics. In: Wu, X.N. (eds) Plant Phosphoproteomics. Methods in Molecular Biology, vol 2358. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1625-3_11
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DOI: https://doi.org/10.1007/978-1-0716-1625-3_11
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