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OxiSelect Total Glutathione (GSSG/GSH) Assay Kit

MBS168487

20 Assays

280 USD

Assay Kit

OxiSelect Total Glutathione (GSSG/GSH) Assay Kit

[OxiSelect Total Glutathione (GSSG/GSH)]

[OxiSelect Total Glutathione (GSSG/GSH) Assay Kit]

Upon receipt, store the NADPH at -80 degree C. Prepare single use aliquots and avoid multiple freeze/thaw cycles. Store the remaining kit components at 4 degree C.

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Oxidative Stress/ Damage; Antioxidant Assays; Glutathione Assay; Glutathione Assay

Principle of the Assay: The OxiSelect Total Glutathione Assay Kit is a quantitative assay for measuring the total glutathione content within a sample (GSH/GSSG). Glutathione Reductase reduces oxidized glutathione (GSSG) to reduced glutathione (GSH) in the presence of NADPH. Subsequently, the chromogen reacts with the thiol group of GSH to produce a colored compound that absorbs at 405 nm (Figure 1). The total glutathione content in unknown samples is determined by comparison with the predetermined glutathione standard curve. The rate of chromophore production is proportional to the concentration of glutathione within the sample. The rate can be determined from the absorbance change over time. Metaphosphoric acid is provided to remove interfering proteins or enzymes from samples. Background/Introduction: Oxidative stress occurs when there is an excess of free radicals, or reactive oxygen species (ROS), in the body. Research has shown that excessive ROS accumulation will lead to cellular injury, such as damage to DNA, proteins, and lipid membranes. ROS damage has been implicated in the development of many physiological problems, such as ageing, asthma, arthritis, diabetes, cancer, inflammation, cardiovascular disease, atherosclerosis, Down's Syndrome, and neurodegenerative diseases. Glutathione is a key intracellular tripeptide thiol composed of glutamic acid, cysteine, and glycine. Glutathione helps protect cells from free radical damage by acting as an antioxidant. Within cells, glutathione exists in reduced (GSH) and oxidized (GSSG) states. In healthy cells and tissue, more than 90% of the total glutathione pool is in the reduced form (GSH) while less than 10% exists in the disulfide form (GSSG). The high GSH concentration is because the enzyme that transitions it from its oxidized state (GSSG), glutathione reductase, is constitutively active and inducible upon oxidative stress. An increased GSSG-to-GSH ratio is considered indicative of oxidative stress. Reduced glutathione's thiol group provides reducing equivalents to other unstable ROS, which in turn becomes unstable itself. This unstable GSH readily reacts with another unstable GSH to form a stable GSSG molecule. This reaction is prevalent since glutathione is present in high concentrations. GSSG is subsequently converted to GSH again by the enzyme glutathione reductase. In addition to its role in oxidative stress, glutathione also helps maintain exogenous antioxidants such as vitamins C and E. Glutathione is involved with the breakdown of peroxides. It has a role in regulating the nitric acid cycle. Glutathione can directly bind many inorganic and organic xenobiotic (foreign chemicals) and carcinogenic compounds, such as the heavy metals mercury and arsenic. It is important to the proper function and maximum effect of the immune system. In addition, glutathione is fundamentally involved with many metabolic and biochemical mechanisms such as protein and prostaglandin synthesis, DNA synthesis and repair, maintenance of disulfie bonds in proteins, enzyme activation and amino acid transport across cell membranes. OxiSelect Total Glutathione Assay Kit is a quantitative assay for measuring the total concentration of glutathione, which encompasses both reduced and oxidized glutathione (GSH and GSSG) from plasma, blood, saliva, urine, tissue extracts, and plant or mammalian cell lysates. The kit employs a simple enzymatic recycling reaction for total glutathione quantification. The kit has a detection sensitivity limit of 8 nM. Each kit provides sufficient reagents to perform up to 100 assays, including standard curve and unknown samples.

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20 Assays

280 USD

100 Assays

505