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Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae.

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

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.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Natriumchlorid, ACS reagent, ≥99.0%
Sigma-Aldrich
Glycerin, for molecular biology, ≥99.0%
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylendiamin, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
Glycin, BioUltra, Molecular Biology, ≥99.0% (NT)
Sigma-Aldrich
Salzsäure, 36.5-38.0%, BioReagent, Molecular Biology
Sigma-Aldrich
Ethylenglykol-bis(2-aminoethylether)-N,N,N′,N′-Tetraessigsäure, Molecular Biology, ≥97.0%
Sigma-Aldrich
Natriumphosphat, BioReagent, Molecular Biology, anhydrous, ≥98%
Sigma-Aldrich
Kaliumcyanid, BioUltra, ≥98.0% (AT)
Supelco
Bicinchoninsäure -Lösung
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Ethylendiamintetraessigsäure Dinatriumsalz Dihydrat, ACS reagent, 99.0-101.0%
Sigma-Aldrich
Bromphenolblau, titration: suitable
Sigma-Aldrich
Nitrotetrazolium-Blauchlorid, ≥90.0% (HPLC)
Sigma-Aldrich
Natriumphosphat, BioXtra, ≥99.0%