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Detection of Ascorbate Peroxidase (APX) Activity in Plant Tissues: Using Non-denaturing PAGE and Spectrophotometric Assay

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ROS Signaling in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2798))

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Abstract

At the cellular level, the generation of reactive oxygen species (ROS), such as hydrogen peroxide (H2O2), due to different abiotic or biotic stress, causes oxidative stress that induces an imbalance in the metabolism. Among the different H2O2-scavenging enzymatic antioxidants, ascorbate peroxidase (APX) is a heme-peroxidase that plays an important role in the ascorbate-glutathione pathway using ascorbate to reduce H2O2 to water. Using non-denaturing polyacrylamide gel electrophoresis (PAGE) in combination with a spectrophotometric assay for APX activity, the protocol allows identifying diverse APX isozymes present in different organs and plant species.

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Acknowledgments

The research is supported by the European Regional Development Fund which cofinanced grants from the Ministry of Science and Innovation (PID2019-103924GB-I00 and CPP2021-008703), Spain. Marta Rodríguez-Ruiz acknowledges a postdoctoral contract (MCIN/AEI/10.13039/501100011033) associated with the grant CPP2021-008703.

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Authors and Affiliations

  1. Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Stress, Development and Signalling in Plants, Estación Experimental del Zaidín (Spanish National Research Council, CSIC), Granada, Spain

    Marta Rodríguez-Ruiz, María A. Muñoz-Vargas & Francisco J. Corpas

  2. Laboratory of Extremophile Plants, Center of Biotechnology of Borj Cedria, Hammam-Lif, Tunisia

    Hayet Houmani

  3. Granada, Spain

    José M. Palma

Authors
  1. Marta Rodríguez-Ruiz
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  2. Hayet Houmani
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  3. María A. Muñoz-Vargas
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  4. José M. Palma
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  5. Francisco J. Corpas
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Corresponding author

Correspondence to Marta Rodríguez-Ruiz .

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Editors and Affiliations

  1. Spanish National Research Council, Estación Experimental del Zaidín, Granada, Granada, Spain

    Francisco J. Corpas

  2. Spanish National Research Council, Estación Experimental del Zaidín, Granada, Spain

    José M. Palma

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Rodríguez-Ruiz, M., Houmani, H., Muñoz-Vargas, M.A., Palma, J.M., Corpas, F.J. (2024). Detection of Ascorbate Peroxidase (APX) Activity in Plant Tissues: Using Non-denaturing PAGE and Spectrophotometric Assay. In: Corpas, F.J., Palma, J.M. (eds) ROS Signaling in Plants . Methods in Molecular Biology, vol 2798. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3826-2_16

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  • DOI: https://doi.org/10.1007/978-1-0716-3826-2_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3825-5

  • Online ISBN: 978-1-0716-3826-2

  • eBook Packages: Springer Protocols

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