Analysis of GSH and GSSG after derivatization with N-ethylmaleimide

  title={Analysis of GSH and GSSG after derivatization with N-ethylmaleimide},
  author={Daniela Giustarini and Isabella Dalle-Donne and A. Milzani and Paolo Fanti and Ranieri Rossi},
  journal={Nature Protocols},
This protocol describes a procedure for determining glutathione (GSH) and glutathione disulfide (GSSG) concentrations in blood and other tissues. Artifactual oxidation to GSSG of 5–15% of the GSH found in a sample can occur during deproteination of biological samples with any of the commonly used acids, with consequent marked overestimation of GSSG. This can be prevented by derivatizing GSH with the alkylating agent N-ethylmaleimide (NEM) to form GS-NEM before acid deproteination, followed by… 

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