Superoxide dismutase isozyme detection using two-dimensional gel electrophoresis zymograms

Journal of Pharmaceutical and Biomedical Analysis
Ploypat NiyomployPolkit Sangvanich

Abstract

Superoxide dismutases (SODs) are ubiquitous antioxidant enzymes involved in cell protection from reactive oxygen species. Their antioxidant activities make them of interest to applied biotechnology industries and are usually sourced from plants. SODs are also involved in stress signaling responses in plants, and can be used as indicators of these responses. In this article, a suitable method for the separation of different SOD isoforms using two-dimensional-gel electrophoresis (2D-GE) zymograms is reported. The method was developed with a SOD standard from bovine erythrocytes and later applied to extracts from Stemona tuberosa. The first (non-denaturing isoelectric focusing) and second (denaturing sodium dodecylsulphate-polyacrylamide gel electrophoresis) dimensions of duplicate 2D-GE gels were stained with either Coomassie brilliant blue G-250 for total protein visualization, or SOD activity (zymogram) using riboflavin/nitroblue tetrazolium. For confirmation, putative SOD activity positive spots were subject to trypsin digestion and nano-liquid chromatography tandem mass spectrometry, followed by searching the MASCOT database for potential identification. The method could separate different SOD isoforms from a plant extract an...Continue Reading

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Citations

Mar 5, 2016·Bioscience, Biotechnology, and Biochemistry·Jutarmas JungsukcharoenPolkit Sangvanich
Mar 11, 2016·PLoS Neglected Tropical Diseases·Diana TamayoOrville Hernández

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