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Updated: May 17, 2026

Rapid Quantification of Oxidized and Reduced Forms of Glutathione Using Ortho -phthalaldehyde in Cultured Mammalian Cells In Vitro
Published on: June 28, 2024
Oxidant-induced regulation of glutathione synthesis
1University Of Alabama At Birmingham, Alabama, USA.
Abstract:
This unit describes protocols for characterizing the expression of two glutathione biosynthesis enzymes: gamma-glutamylcysteine synthase (GCS) and gamma-glutamyl transpeptidase (GGT) in response to oxidants. GCS catalyzes the first and rate-limiting step of glutathione synthesis, while GGT degrades extracellular glutathione (GSH) to provide the amino acids required for intracellular synthesis of GSH. Northern blot hybridization is used to quantitatively assess the mRNAs for the two enzymes in response to oxidant. Nuclear run-on is used to determine the rate of transcription.
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