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Determination of Reduced and Total Glutathione Content in Extremophilic Microalga Galdieria phlegrea.

Bio-protocol (2017-07-05)
Giovanna Salbitani, Claudia Bottone, Simona Carfagna
ABSTRACT

Glutathione is an important molecule involved in the primary and secondary metabolism of all organisms. The Glutathione redox status is an indicator of the cellular redox state. Therefore, it is important to have precise methods on hand to determine the glutathione redox status in the cell. In this protocol, we describe an improved spectrophotometric method to estimate the content of reduced (GSH) and oxidized (GSSG) forms of glutathione in the extremophilic microalga Galdieria phlegrea.

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Sigma-Aldrich
Acido etilendiamminotetraacetico, suitable for electrophoresis, Molecular Biology, 99.0-101.0% (titration)
Sigma-Aldrich
L-Glutathione reduced, suitable for cell culture, BioReagent, ≥98.0%, powder
Sigma-Aldrich
5,5′-ditiobis(acido 2-nitrobenzoico), ≥98%, BioReagent, suitable for determination of sulfhydryl groups
Sigma-Aldrich
Glutatione reduttasi, ammonium sulfate suspension, 100-300 units/mg protein (biuret)
Sigma-Aldrich
Fosfato di sodio, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium phosphate monobasic, BioXtra, ≥99.0%
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide phosphate, reduced tetra(cyclohexylammonium) salt, ≥93%