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US 2012/0058501 A1Published application
Oxidized glutathione assay
AssigneePromega Corporation
InventorsFen Huang, Dieter Klaubert, John Shultz, Wenhui Zhou
Life of this patent
Estimating the term…
Abstract
The present invention provides an assay for detection of oxidized glutathione (GSSG).
Claims41
A method for detecting GSSG comprising:
- a. contacting a sample with a sulfhydryl modifying agent;
- b. contacting the sample with an excess of a reducing agent, glutathione-S-transferase, and a substrate which is converted to a signal-generating compound in the presence of GSH and glutathione-S-transferase;
- and
- c. detecting a signal.
1 more element
A method for determining GSH:GSSG ratio comprising:
- a. removing a portion of a sample;
- b. contacting the remaining sample with a sulfhydryl modifying agent;
- c. adding to the remaining sample an excess of a reducing agent, glutathione-S-transferase, and a substrate which is converted to a signal-generating compound in the presence of GSH and glutathione-S-transferase to form a first solution;
- d. measuring the signal of the first solution;
- e. contacting the removed sample with glutathione-S-transferase and a substrate which is converted to a signal-generating compound in the presence of GSH and glutathione-S-transferase to form a second solution;
- f. measuring the signal of the second solution;
- and
- g. comparing the signal of the second solution to the signal of the first solution to determine the ratio of GSH to GSSG.
5 more elements
A method of determining oxidative stress in a sample comprising:
- a. contacting the sample with a sulfhydryl modifying agent;
- b. contacting the sample with an excess of a reducing agent, glutathione-S-transferase, and a substrate which is converted to a signal-generating compound in the presence of GSH and glutathione-S-transferase;
- and
- c. measuring the signal.
1 more element
Glutathione-S-transferase that is essentially free of GSH.
Dependencies are inferred from the claim text.