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  • Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae.

Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae.

Bio-protocol (2021-03-05)
Louise Thines, Pierre Morsomme
摘要

Superoxide dismutases (SODs) act as a primary defence against reactive oxygen species (ROS) by converting superoxide anion radicals (O2 -) into molecular oxygen (O2) and hydrogen peroxide (H2O2). Members of this enzyme family include CuZnSODs, MnSODs, FeSODs, and NiSODs, depending on the nature of the cofactor that is required for proper activity. Most eukaryotes, including yeast, possess CuZnSOD and MnSOD. This protocol aims at assessing the activity of the yeast Saccharomyces cerevisiae MnSOD Sod2p from cellular extracts using nitroblue tetrazolium staining. This method can be used to estimate the cellular bioavailability of Mn2+ as well as to evaluate the redox state of the cell.

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氯化钠, ACS reagent, ≥99.0%
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甘油, for molecular biology, ≥99.0%
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Triton X-100, laboratory grade
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四甲基乙二胺, BioReagent, suitable for electrophoresis, ≥99.0%
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甘氨酸, BioUltra, for molecular biology, ≥99.0% (NT)
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盐酸, 36.5-38.0%, BioReagent, for molecular biology
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乙二醇-双(2-氨基乙醚)-N,N,N′,N′-四乙酸, for molecular biology, ≥97.0%
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乙二胺四乙酸 二钠盐 二水合物, ACS reagent, 99.0-101.0%
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磷酸钠 一元, BioReagent, for molecular biology, anhydrous, ≥98%
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二喹啉甲酸 溶液
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磷酸钠 二元, BioXtra, ≥99.0%
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氯化四唑氮蓝, ≥90.0% (HPLC)
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溴酚蓝, titration: suitable