Preparation for oxidative stress (POS), i.e., the buildup of endogenous antioxidants during metabolic depression or low oxygen stress conditions, has been observed in at least 8 animal phyla under controlled conditions in laboratory. Despite the expected implications on the endurance to extreme environments and ecosystem occupation, the extent to which POS occurs in animals under natural conditions remains unexplored. Therefore, we took advantage of the natural history of the Brazilian Caatinga's frog Proceratophrys cristiceps to investigate the modulation of endogenous antioxidants and redox balance in their skeletal muscle and to verify if POS occurs under natural conditions. Expectedly, estivating frogs had low levels of the oxidative metabolism enzymes. Citrate synthase and isocitrate dehydrogenase activities were 36% and 25% lower than those in active frogs respectively. We found an overall upregulation of antioxidants in estivating P. cristiceps. Reduced glutathione concentration was 61% higher in estivating frogs than that in active animals. During estivation the activities of the hydroperoxide detoxification enzymes catalase, glutathione peroxidase, and glutathione HO-peroxidase were 48%, 57%, and 78% greater than those during the rainy season. Moreover, estivating frogs had a 47% lower ratio of disulfide to total glutathione levels than active frogs. Our findings confirm the occurrence of 'preparation for oxidative stress' in naturally estivating frogs and paves the way for further research on the redox biology of animals under natural settings. Such approach might reveal biochemical strategies under ecologically relevant scenarios.