I have to quantify the enzymatic activities of SOD in plants using a method that employs hypoxanthine-xanthine oxidase to create O2. The colorimetric reaction was performed by combining alpha-naphthylamine and sulfanilic acid. I measured the absorbance at 550 nm and recorded the absorbance, including a blank without the plant extract to determine the maximum production of O2, as well as the absorbance of the plant extract.
Based on this information how can i calculate activities of the SOD?
To convert from absorbance to product concentration, there are two methods.
1. Standard curve. Prepare an evenly-spaced set of concentrations of product under identical conditions as the enzyme reaction (except no enzyme), measure their absorbances, plot absorbance versus concentration. Read off product concentrations from this curve.
2. Known extinction coefficient. If there is a published value of the extinction coefficient difference between product and substrate, you can use this value and Beer's law to calculate product concentration in the enzyme assay.
Sure, you can follow the enzymatic reaction spectrophotometrically.
https://www.frontiersin.org/journals/behavioral-neuroscience/articles/10.3389/fnbeh.2023.1304006/full
please check my publication
https://www.sciencedirect.com/science/article/abs/pii/S0141813014000026
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