GSH+GSSG / GSH Assay Kit (Colorimetric) (ab239709)
Key features and details
- Detection method: Colorimetric
- Platform: Microplate reader
Overview
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Product name
GSH+GSSG / GSH Assay Kit (Colorimetric)
See all Glutathione kits -
Detection method
Colorimetric -
Product overview
GSH+GSSG / GSH Assay Kit (Colorimetric) (ab239709) provides a convenient, colorimetric method for analyzing either total glutathione or the reduced form glutathione alone using a microtiter plate reader.
The assay is based on the glutathione recycling system by DTNB and glutathione reductase. DTNB and glutathione (GSH) react to generate 2-nitro-5-thiobenzoic acid which has a yellow color. Therefore, GSH concentration can be determined by measuring absorbance at 412 nm.
The generated GSSG can be reduced back to GSH by glutathione reductase, and GSH reacts with DTNB again to produce more 2-nitro-5-thiobenzoic acid. Therefore, the recycling system dramatically improves the sensitivity of total glutathione detection.
The kit includes the 5-Sulfosalicylic acid (SSA) for the removal of proteins from samples and for the protection of GSH oxidation and g-glutamyl transpeptidase reaction.
The kit can quantify glutathione from 1-100 ng/well in a 200 mL reaction. For detecting lower glutathione concentrations, such as in blood samples, increasing reaction time will generate stronger signal.
The kit can also specifically detect the reduced form of glutathione (GSH) by omitting the glutathione reductase from the reaction mixture. The sensitivity for detecting the reduced form of glutathione (without recycling system) is 100 times lower than detecting the total glutathione.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Glutathione Reaction Buffer 1 x 100ml Glutathione Reductase 2 x 25µl Glutathione Substrate (DTNB) 2 vials GSH Standard (lyophilized, MW 307) 2 x 1mg NADPH Generating Mix (lyophilized) 2 vials Sulfosalicylic Acid (SSA, 1 gram) 1 unit -
Research areas
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Relevance
Glutathione is a small peptide composed of three amino acids: cysteine, glutamic acid, and glycine and is present in tissues in concentrations as high as one millimolar. It contains an unusual peptide linkage between the amine group of cysteine and the carboxyl group of the glutamate side chain. Glutathione is involved in detoxification, it binds to toxins, such as heavy metals, solvents, and pesticides, and transforms them into a form that can be excreted in urine or bile. It is also an important antioxidant, helping to maintain the -SH groups of proteins in their reduced form. Chronic functional glutathione deficiency is associated with glucose 6-phosphate dehydrogenase deficiency, immune disorders, an increased incidence of malignancies, and in the case of HIV disease, probably accelerated pathogenesis of the disease. Acute manifestations of functional glutathione deficiency can be seen in those who have taken an overdosage of acetaminophen (paracetamol). This results in depletion of glutathione in the hepatocytes, leading to liver failure and death. -
Alternative names
- Glutathione
- GSH
- Oxidised glutathione
- Reduced glutathione
Images
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Glutathione Standard Curve.
Various amounts of standard glutathione was added to the glutathione reaction and incubated for 10 min according to the kit instructions. Absorbance was measured at O.D. 405 nm.
