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

D8130

Sigma-Aldrich

5,5′-Dithiobis(2-nitrobenzoic acid), ≥98%, BioReagent, suitable for determination of sulfhydryl groups

N5130

Sigma-Aldrich

β-Nicotinamide adenine dinucleotide phosphate, reduced tetra(cyclohexylammonium) salt, ≥93%

E5134

Sigma-Aldrich

Ethylenediaminetetraacetic acid disodium salt dihydrate, suitable for electrophoresis, for molecular biology, 99.0-101.0% (titration)

S7907

Sigma-Aldrich

Sodium phosphate dibasic, BioXtra, ≥99.0%

G6013

Sigma-Aldrich

L-Glutathione reduced, suitable for cell culture, BioReagent, ≥98.0%, powder

S8282

Sigma-Aldrich

Sodium phosphate monobasic, BioXtra, ≥99.0%

G3664

Sigma-Aldrich

Glutathione Reductase from baker′s yeast (S. cerevisiae), ammonium sulfate suspension, 100-300 units/mg protein (biuret)