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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
Referencia del producto
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Sigma-Aldrich
Cloruro de sodio, ACS reagent, ≥99.0%
Sigma-Aldrich
Glicerol, for molecular biology, ≥99.0%
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N,N,N′,N′-Tetrametiletilendiamina, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Glicina, BioUltra, for molecular biology, ≥99.0% (NT)
Sigma-Aldrich
Ácido clorhídrico, 36.5-38.0%, BioReagent, for molecular biology
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, for molecular biology, ≥97.0%
Sigma-Aldrich
Ácido etilenodiaminotetraacético disodium salt dihydrate, ACS reagent, 99.0-101.0%
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Sodium phosphate monobasic, BioReagent, for molecular biology, anhydrous, ≥98%
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Potassium cyanide, BioUltra, ≥98.0% (AT)
Supelco
Ácido bicinconínico solution
Sigma-Aldrich
Fosfato de sodio dibasic, BioXtra, ≥99.0%
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Nitroazul de tetrazolio, ≥90.0% (HPLC)
Sigma-Aldrich
Bromophenol Blue, titration: suitable