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

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
5,5′-Dithiobis(2-nitrobenzoic acid), ≥98%, BioReagent, suitable for determination of sulfhydryl groups
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide phosphate, reduced tetra(cyclohexylammonium) salt, ≥93%
Sigma-Aldrich
Ethylenediaminetetraacetic acid disodium salt dihydrate, suitable for electrophoresis, for molecular biology, 99.0-101.0% (titration)
Sigma-Aldrich
L-Glutathione reduced, suitable for cell culture, BioReagent, ≥98.0%, powder
Sigma-Aldrich
Glutathione Reductase from baker′s yeast (S. cerevisiae), ammonium sulfate suspension, 100-300 units/mg protein (biuret)
Sigma-Aldrich
Sodium phosphate dibasic, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium phosphate monobasic, BioXtra, ≥99.0%