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

Autor: Rodríguez-Ruiz M; 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.rodriguez@eez.csic.es., Houmani H; Laboratory of Extremophile Plants, Center of Biotechnology of Borj Cedria, Hammam-Lif, Tunisia., Muñoz-Vargas MA; 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., Palma JM; Spanish National Research Council, Estación Experimental del Zaidín, Granada, Spain., Corpas FJ; 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.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2798, pp. 223-234.
DOI: 10.1007/978-1-0716-3826-2_16
Abstrakt: At the cellular level, the generation of reactive oxygen species (ROS), such as hydrogen peroxide (H 2 O 2 ), due to different abiotic or biotic stress, causes oxidative stress that induces an imbalance in the metabolism. Among the different H 2 O 2 -scavenging enzymatic antioxidants, ascorbate peroxidase (APX) is a heme-peroxidase that plays an important role in the ascorbate-glutathione pathway using ascorbate to reduce H 2 O 2 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.
(© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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