Determining superoxide dismutase content and catalase activity in mammalian cell lines
Ilzé Engelbrecht,
Suranie Horn,
John P. Giesy,
Rialet Pieters
Affiliations
Ilzé Engelbrecht
Unit for Environmental Sciences and Management, North-West University, Potchefstroom, 2520, South Africa; Occupational Hygiene and Health Research Initiative, North-West University, Potchefstroom, 2520, South Africa; Corresponding author at: Unit for Environmental Sciences and Management, North-West University, Potchefstroom 2520, South Africa.
Suranie Horn
Unit for Environmental Sciences and Management, North-West University, Potchefstroom, 2520, South Africa; Occupational Hygiene and Health Research Initiative, North-West University, Potchefstroom, 2520, South Africa
John P. Giesy
Toxicology Centre, University of Saskatchewan, Saskatoon, Canada; Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada; Department of Integrative Biology and Center for Integrative Toxicology, Michigan State University, East Lansing, MI 48824, USA; Department of Environmental Science, Baylor University, One Bear Place #97266, Waco, TX 76798, USA
Rialet Pieters
Unit for Environmental Sciences and Management, North-West University, Potchefstroom, 2520, South Africa
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).
