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Reactive oxygen species (ROS) production during immune responses is tightly regulated by the multiple phosphorylation of plasma membrane-localized NADPH oxidases, respiratory burst oxidase homologs (RBOHs). It is known that calcium-dependent protein kinases (CDPKs) and other kinases phosphorylate N-terminal region of RBOH and activate ROS production during immune responses. In this chapter, we describe protocols for the preparation of anti-phosphopeptide antibodies, and the detection of phosphorylated RBOH during immune responses using anti-phospho-RBOH antibody in Nicotiana benthamiana.

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Detection of Phosphorylated NADPH Oxidase by Antibody During Plant Immune Responses

  • 实验方案

  • 发表日期：

    2026年

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  • Yuta Hino,
  • Hirofumi Yoshioka

摘要

Reactive oxygen species (ROS) production during immune responses is tightly regulated by the multiple phosphorylation of plasma membrane-localized NADPH oxidases, respiratory burst oxidase homologs (RBOHs). It is known that calcium-dependent protein kinases (CDPKs) and other kinases phosphorylate N-terminal region of RBOH and activate ROS production during immune responses. In this chapter, we describe protocols for the preparation of anti-phosphopeptide antibodies, and the detection of phosphorylated RBOH during immune responses using anti-phospho-RBOH antibody in Nicotiana benthamiana.