Glutathione (GSH) is the highest concentration non-protein thiol in mammalian cells and is present in concentrations of 0.5 to 10 mM. GSH plays a key role in many biological processes, including the synthesis of proteins and DNA, the transport of amino acids, and the protection of cells against oxidation. Harmful hydrogen peroxide cellular levels are minimized by the enzyme glutathione peroxidase (GP) using GSH as a reductant. The oxidized GSH dimer, GSSG, is formed from GSH and peroxide by the GP reaction. An important role of GSSG in the NFkB activating signal cascade is suggested by the fact that the potent NFkB inducer TPA increases intracellular GSSG levels and GSSG/GSH ratios.
The Glutathione Kit is colorimetric assay designed to quantitatively measure glutathione (GSH), and oxidized glutathione (GSSG) present in a variety of samples. No separation or washing is required. Total glutathione is the sum of GSSG plus GSH. A liquid GSSG standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. The kit utilizes a colorimetric substrate that reacts with the free thiol group on GSH to yield a highly colored product. All reagents are in solution and require simple dilution
for use in the assay. Free GSH concentration in the sample is calculated from the difference between the total GSH determined and the GSH generated from GSSG. The concentration of GSH can be determined either as an endpoint read of the color developed at 405 nm or by measuring the rate of color development at 405 nm.
Blood, serum, plasma, urine, erythrocytes and cell lysate samples have been validated with this assay.
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