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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
초록

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.

MATERIALS
제품 번호
브랜드
제품 설명

Sigma-Aldrich
Bromophenol Blue, titration: suitable
Sigma-Aldrich
Sodium phosphate dibasic, BioXtra, ≥99.0%
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylenediamine, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
Sodium phosphate monobasic, BioReagent, Molecular Biology, anhydrous, ≥98%
Sigma-Aldrich
Glycerol, for molecular biology, ≥99.0%
Sigma-Aldrich
Triton X-100, laboratory grade
Supelco
Bicinchoninic Acid solution
Sigma-Aldrich
Ethylenediaminetetraacetic acid disodium salt dihydrate, ACS reagent, 99.0-101.0%
Sigma-Aldrich
Potassium cyanide, BioUltra, ≥98.0% (AT)
Sigma-Aldrich
Hydrochloric acid, 36.5-38.0%, BioReagent, Molecular Biology
Sigma-Aldrich
Nitrotetrazolium Blue chloride, ≥90.0% (HPLC)
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, Molecular Biology, ≥97.0%
Sigma-Aldrich
Glycine, BioUltra, Molecular Biology, ≥99.0% (NT)
Sigma-Aldrich
Sodium chloride, ACS reagent, ≥99.0%