Determining superoxide dismutase content and catalase activity in mammalian cell lines

Traditional methods for determining superoxide dismutase (SOD) content and catalase (CAT) activity rely on measuring the absorbance of individual tissue (biological) samples using a cuvette and spectrophotometer, rather than cell cultures. Although there are kits available for SOD and CAT assays, these allow for high-throughput analysis of samples and might be too expensive for research laboratories in countries from the Global South, such as South Africa. This paper describes a simple and cost-effective method to determine SOD content and CAT activity in mammalian cell cultures following exposure to environmental chemical mixtures by measuring absorbance in 96-well microplates. Moreover, the equipment used for this method is considered standard for cell culture laboratories, while the reagents and consumables are easily obtainable. • Antioxidant enzyme levels can be measured in vitro in cell cultures. • The supernatant obtained can be used to determine protein concentration, SOD content, and CAT activity. • This method is simple and affordable, allowing for the analysis of multiple samples (up to 32 samples per microplate).