Abstract
We provide an optimized protocol for a double staining technique to analyze superoxide dismutase enzymatic isoforms Cu-Zn SOD (Sod1) and Mn-SOD (Sod2) and catalase in the same polyacrylamide gel. The use of NaCN, which specifically inhibits yeast Sod1 isoform, allows the analysis of Sod2 isoform while the use of H2O2 allows the analysis of catalase. The identification of a different zymography profiling of SOD and catalase isoforms in different yeast species allowed us to propose this technique as a novel yeast identification and classification strategy.
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Acknowledgements
This work has been supported by grants AGL2011-24353 and AGL2014-52985-R from the Spanish Ministry of Economy and Competitiveness (MINECO) to E.M., and it has been performed within the Program VLC/Campus, Microcluster IViSoCa (Innovation for a Sustainable Viticulture and Quality), and Microcluster BBLM (Model Yeasts in Biomedicine & Biotechnology). E.G.-S. was a predoctoral fellow of the JAE program from the CSIC (Spanish National Research Council). R.G.-P. was a postdoctoral researcher at Universitat de València.
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Gamero-Sandemetrio, E., Gómez-Pastor, R., Matallana, E. (2017). Zymography Methods to Simultaneously Analyze Superoxide Dismutase and Catalase Activities: Novel Application for Yeast Species Identification. In: Wilkesman, J., Kurz, L. (eds) Zymography. Methods in Molecular Biology, vol 1626. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7111-4_17
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DOI: https://doi.org/10.1007/978-1-4939-7111-4_17
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