ArticleMethods in molecular biology (Clifton, N.J.)2024
Determination of ROS Generated by Arabidopsis Xanthine Dehydrogenase1 (AtXDH1) Using Nitroblue Tetrazolium (NBT) and 3,3'-Diaminobenzidine (DAP).
Article in Methods in molecular biology (Clifton, N.J.), 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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7 citing papers in PubMed.
- Functional Characterization ofPlants (Basel, Switzerland) · 2026Article
- Plant-Specific PB1 Domain Complex Mediates Poplar Defence and Is Targeted by Rust Fungus Secreted Protein MlpSD1.Molecular plant pathology · 2026Article
- Article
- VdEGe1, a cell wall-degrading enzyme gene from Verticillium dahliae required for cotton pathogenesis.BMC microbiology · 2026Article
- Iron status modulates cadmium tolerance: mechanistic insights fromFrontiers in plant science · 2026Article
- Maize 4-coumarate coenzyme A ligaseGM crops & food · 2025Article
- The Plant Growth-Promoting BacteriumMicroorganisms · 2025Article
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Abstract
Plants generate reactive oxygen species (ROS) during different metabolic processes, which play an essential role in coordinating growth and response. ROS levels are sensitive to environmental stresses and are often used as a marker for stress in plants. While various methods can detect ROS changes, histochemical staining with nitroblue tetrazolium (NBT) and 3,3'-diaminobenzidine (DAB) is a popular method, though it has faced criticism. This staining method is advantageous as it enables both the quantification and localization of ROS and the identification of the enzymatic origin of ROS in plants, cellular compartments, or gels. In this protocol, we describe the use of NBT and DAP staining to detect ROS generation under different stresses such as nitrogen starvation, wounding, or UV-C. Additionally, we describe the use of NBT staining for detecting enzymatic generation of ROS in native and native SDS PAGE gels. Our protocol also outlines the separation and comparison of the origin of ROS generated by xanthine dehydrogenase1 (XDH1) using different substrates.
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