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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
要旨

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
製品番号
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製品内容

Sigma-Aldrich
塩化ナトリウム, ACS reagent, ≥99.0%
Sigma-Aldrich
グリセロール, for molecular biology, ≥99.0%
Sigma-Aldrich
N,N,N′,N′-テトラメチルエチレンジアミン, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
グリシン, BioUltra, Molecular Biology, ≥99.0% (NT)
Sigma-Aldrich
塩酸, 36.5-38.0%, BioReagent, Molecular Biology
Sigma-Aldrich
エチレングリコール-ビス(2-アミノエチルエーテル)-N,N,N′,N′-四酢酸, Molecular Biology, ≥97.0%
Sigma-Aldrich
リン酸ナトリウム 一塩基性, BioReagent, Molecular Biology, anhydrous, ≥98%
Sigma-Aldrich
シアン化カリウム, BioUltra, ≥98.0% (AT)
Supelco
ビシンコニン酸 溶液
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
エチレンジアミン四酢酸 二ナトリウム塩 二水和物, ACS reagent, 99.0-101.0%
Sigma-Aldrich
ブロモフェノールブルー, titration: suitable
Sigma-Aldrich
ニトロテトラゾリウムブルークロリド, ≥90.0% (HPLC)
Sigma-Aldrich
リン酸ナトリウム 二塩基性, BioXtra, ≥99.0%