Redox homeostasis is crucial for cell function, and, in eukaryotic cells, studying it in a compartmentalized way is essential due to the redox variations between different organelles. The redox state of organelles is largely determined by the redox potential of glutathione, E(GSH), and the concentration of its reduced and oxidized species, [GS]. The Golgi apparatus is an essential component of the secretory pathway, yet little is known about the concentration or redox state of GSH in this organelle. Here, we characterized the redox state of GSH in the Golgi apparatus using a combination of microscopy and proteomics methods. Our results prove that the Golgi apparatus is a highly oxidizing organelle with a strikingly low GSH concentration (E(GSH) = - 157 mV, 1-5 mM). These results fill an important gap in our knowledge of redox homeostasis in subcellular organelles. Moreover, the new Golgi-targeted GSH sensors allow us to observe dynamic changes in the GSH redox state in the organelle and pave the way for further characterization of the Golgi redox state under various physiological and pathological conditions.