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Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae.

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

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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Description du produit

Sigma-Aldrich
Chlorure de sodium, ACS reagent, ≥99.0%
Sigma-Aldrich
Glycérol, for molecular biology, ≥99.0%
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylenediamine, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
Glycine, BioUltra, for molecular biology, ≥99.0% (NT)
Sigma-Aldrich
Acide chlorhydrique, 36.5-38.0%, BioReagent, for molecular biology
Sigma-Aldrich
Acide éthylène glycol-bis(2-aminoéthyléther)-N,N,N′,N′-tétraacétique, for molecular biology, ≥97.0%
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Phosphate de sodium monobasic, BioReagent, for molecular biology, anhydrous, ≥98%
Sigma-Aldrich
Potassium cyanide, BioUltra, ≥98.0% (AT)
Sigma-Aldrich
Acide éthylènediaminetétraacétique disodium salt dihydrate, ACS reagent, 99.0-101.0%
Supelco
Acide bicinchoninique solution
Sigma-Aldrich
Bromophenol Blue, titration: suitable
Sigma-Aldrich
Phosphate de sodium dibasic, BioXtra, ≥99.0%
Sigma-Aldrich
Chlorure de bleu de nitrotétrazolium, ≥90.0% (HPLC)