Passa al contenuto
Merck

Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae.

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

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.

MATERIALI
Numero di prodotto
Marchio
Descrizione del prodotto

Sigma-Aldrich
Sodio cloruro, ACS reagent, ≥99.0%
Sigma-Aldrich
Glicerolo, Molecular Biology, ≥99.0%
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
N,N,N′,N′-tetrametil-etilendiammina, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
Acido etilendiamminotetraacetico, ACS reagent, 99.0-101.0%
Sigma-Aldrich
Acido tetraacetico etilenico -bis(2-aminoetiletere)-N,N,N′,N′, Molecular Biology, ≥97.0%
Sigma-Aldrich
Sodium phosphate monobasic, BioReagent, Molecular Biology, anhydrous, ≥98%
Sigma-Aldrich
Glicina, BioUltra, Molecular Biology, ≥99.0% (NT)
Sigma-Aldrich
Acido cloridrico, 36.5-38.0%, BioReagent, Molecular Biology
Sigma-Aldrich
Potassium cyanide, BioUltra, ≥98.0% (AT)
Supelco
Acido bicinconinico
Sigma-Aldrich
Fosfato di sodio, BioXtra, ≥99.0%
Sigma-Aldrich
Blu nitrotetrazolo cloruro, ≥90.0% (HPLC)
Sigma-Aldrich
Bromophenol Blue, titration: suitable