The endogenous generation of reactive oxygen species (ROS), previously perceived as a detrimental by-product of cellular processes, is now recognized as a critical component of intracellular signaling. Exploration of these biological signaling functions requires understanding the complex redox biochemistry and recognizing the compartment-specific elements of ROS generation. The endosomal compartment is increasingly recognized as a source for NADPH oxidase (NOX)-generated signaling ROS. Despite this growing understanding, there are significant limitations to the available detection and measurement systems for endogenous ROS. This chapter provides information about specific methodologies and redox-sensitive probes to guide the investigator and define the critical limitations for many of the available approaches. Although measurement continues to be challenging, the rapid growth and development of new detection systems suggests that our capacity to assign specific signaling roles to endosomal ROS will expand markedly in the next several years.
Keywords: Endosome; Intracellular signaling; NADPH oxidase; Reactive oxygen species.
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