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Merck

Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae.

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

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.

MATERIALES
Número de producto
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Descripción del producto

Sigma-Aldrich
Cloruro de sodio, ACS reagent, ≥99.0%
Sigma-Aldrich
Glicerol, Molecular Biology, ≥99.0%
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
N,N,N′,N′-Tetrametiletilendiamina, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
Ácido etilenodiaminotetraacético disodium salt dihydrate, ACS reagent, 99.0-101.0%
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, 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
Ácido clorhídrico, 36.5-38.0%, BioReagent, Molecular Biology
Sigma-Aldrich
Potassium cyanide, BioUltra, ≥98.0% (AT)
Supelco
Ácido bicinconínico solution
Sigma-Aldrich
Fosfato de sodio dibasic, BioXtra, ≥99.0%
Sigma-Aldrich
Nitroazul de tetrazolio, ≥90.0% (HPLC)
Sigma-Aldrich
Bromophenol Blue, titration: suitable