Abstract
Substantial evidence implicates important roles for both protein phosphorylation and protein degradation in regulation of plant defense responses. Therefore, interest is growing in applying proteomics techniques to investigate these posttranscriptional changes. We have found, however, that most proteins of interest are not visible on two-dimensional (2D) gels without previous prefractionation. This chapter describes the use of preparative denaturing isoelectric focusing to enrich for proteins of specific isoelectric points before separation by 2D gels. This method significantly increases the sensitivity of 2D gelbased comparisons.
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References
Nuhse, T. S., Boller, T., and Peck, S. C. (2003) A plasma membrane syntaxin is phosphorylated in response to the bacterial elicitor flagellin. J. Biol. Chem. 278, 45,248–45,254.
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Serna-Sanz, A., Rairdan, G., Peck, S.C. (2007). Preparative Denaturing Isoelectric Focusing for Enhancing Sensitivity of Proteomic Studies. In: Ronald, P.C. (eds) Plant-Pathogen Interactions. Methods in Molecular Biology, vol 354. Humana Press. https://doi.org/10.1385/1-59259-966-4:99
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DOI: https://doi.org/10.1385/1-59259-966-4:99
Publisher Name: Humana Press
Print ISBN: 978-1-58829-448-7
Online ISBN: 978-1-59259-966-0
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