Engelbrecht Ilzé, Horn Suranie, Giesy John P, Pieters Rialet
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.
MethodsX. 2023 Sep 23;11:102395. doi: 10.1016/j.mex.2023.102395. eCollection 2023 Dec.
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 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).
传统的测定超氧化物歧化酶(SOD)含量和过氧化氢酶(CAT)活性的方法依赖于使用比色皿和分光光度计测量单个组织(生物)样本的吸光度,而非细胞培养物。尽管有用于SOD和CAT检测的试剂盒,但这些试剂盒可实现样本的高通量分析,对于来自全球南方国家(如南非)的研究实验室来说可能过于昂贵。本文描述了一种简单且经济高效的方法,通过测量96孔微孔板中的吸光度来测定哺乳动物细胞培养物在暴露于环境化学混合物后SOD的含量和CAT的活性。此外,该方法所使用的设备在细胞培养实验室中被视为标准设备,而试剂和耗材也很容易获得。
• 可以在细胞培养物中测量抗氧化酶水平。
• 获得的上清液可用于测定蛋白质浓度、SOD含量和CAT活性。
• 该方法简单且经济实惠,可对多个样本进行分析(每个微孔板最多32个样本)。