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Analysis of MAPK Activities Using MAPK-Specific Antibodies

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Plant MAP Kinases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1171))

Abstract

Phosphorylation of proteins by mitogen-activated protein kinases is central to many cellular processes, including signal transduction after stress encounter. Thus, assays to identify or characterize MAP kinase activities are a key tool for research in this area. While in-gel kinase assays using isotope-labeled ATP are a powerful tool to investigate the general induction of MAPK activities in any organism, alternative methods using phospho-specific MAPK antibodies are now being established for many model organisms. However, both in-gel kinase assay and phospho-specific western blot analysis do not allow for the unambiguous identification of the activated MAPK. To obtain specificity, initial immunoprecipitation purification of the kinase of interest prior to further analysis can be performed.

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Acknowledgements

We thank the Deutsche Forschungsgemeinschaft (SFB 766 and BR3875/1-1 as part of the ERA-PG PRR CROP consortium) for support to R.W. and A.A.G.

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Authors and Affiliations

  1. Department of Plant Biochemistry, ZMBP, University of Tübingen, Auf der Morgenstelle 32, 72076, Tübingen, Germany

    Roland Willmann, Daniel J. Haischer & Andrea A. Gust

Authors
  1. Roland Willmann
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  2. Daniel J. Haischer
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  3. Andrea A. Gust
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Corresponding author

Correspondence to Andrea A. Gust .

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Editors and Affiliations

  1. Palacky University in Olomouc, Olomouc, Czech Republic

    George Komis

  2. Faculty of Science, Palacky University in Olomouc, Olomouc, Czech Republic

    Jozef Šamaj

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Willmann, R., Haischer, D.J., Gust, A.A. (2014). Analysis of MAPK Activities Using MAPK-Specific Antibodies. In: Komis, G., Šamaj, J. (eds) Plant MAP Kinases. Methods in Molecular Biology, vol 1171. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0922-3_3

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  • DOI: https://doi.org/10.1007/978-1-4939-0922-3_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0921-6

  • Online ISBN: 978-1-4939-0922-3

  • eBook Packages: Springer Protocols

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