Total Glutathione Quantification Kit

Glutathione (GSH) is the most abundant thiol compound in animal tissues, plant tissues, bacteria, and yeast. GSH has many different roles, including protection against reactive oxygen species and the maintenance of protein thiol groups. During these processes, GSH is converted into its oxidized form, glutathione disulfide (GSSG). Since GSSG is then enzymatically reduced by glutathione reductase, GSH is the dominant form in organisms. DTNB (5,5'-Dithiobis(2-nitrobenzoic acid)), known as Ellman’s Reagent, was developed to detect thiol compounds. In 1985, Dr. M. E. Anderson suggested that the glutathione recycling system involving DTNB and glutathione reductase could be used as a highly sensitive glutathione detection method. DTNB and GSH react to generate 5-Mercapto-2-nitrobenzoic acid (TNB). Since TNB is yellow, the GSH concentration in a sample solution can be determined by O.D. measurement at 412 nm absorbance. GSH is regenerated from GS-TNB by glutathione reductase and will again react with DTNB to produce TNB. This recycling reaction improves the sensitivity of total glutathione detection. Total Glutathione Quantification Kit contains all of the necessary reagents for total glutathione measurement, except for those used in sample preparation. 5-Sulfosalicylic acid is recommended for the removal of proteins from sample solutions and for the prevention of GSH oxidation and γ-glutamyl transpeptidase reactions. However, the optimum method for sample preparation differs from sample to sample, so please review the references. This kit can be used to quantify total glutathione concentrations from 1 μM to 100 μM using the standard method. For lower glutathione concentrations, such as in blood samples, longer incubation times are required.

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