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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.

材料
Product Number
品牌
产品描述

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