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Oxidative stress results from excessive accumulation of reactive oxygen species (ROS) and plays a central role in numerous physiological and pathological processes. Accurate quantification of antioxidant enzyme activities is therefore essential in redox biology research. However, data analysis for commonly used assays, such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), is frequently performed using spreadsheets or manual calculations, which are time-consuming and prone to error. Here, we present Redoxyme, a free, open-source, Python-based graphical user interface designed to standardize and automate the calculation of antioxidant enzyme activities. The software integrates protein normalization, enzyme-specific calculation routines, data visualization, and Excel export within an intuitive interface that does not require programming expertise. Redoxyme was validated using experimental data obtained from animal tissues (rats and mice), demonstrating excellent agreement with manual calculations and established analytical methods. Redoxyme provides a practical solution for improving reproducibility and efficiency in antioxidant enzyme activity analysis. The software is currently distributed as a standalone executable for Windows (locally installed), and an interactive web-based calculator implemented in Streamlit, enabling direct use without local installation. The source code and version-controlled development history are openly accessible via GitHub, promoting transparency, reproducibility, community-driven improvements, and can, in principle, be adapted for other operating systems.