Glutathione Assay Kit (Fluorometric) (ab65322)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 2 hr
- Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
Overview
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Product name
Glutathione Assay Kit (Fluorometric)
See all Glutathione kits -
Detection method
Fluorescent -
Sample type
Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media -
Assay type
Quantitative -
Assay time
2h 00m -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
Glutathione Assay Kit (Fluorometric) (ab65322) provides a simple in vitro assay for detection of total glutathione changes during cellular response to toxicity, apoptosis and other conditions. The assay uses the dye monochlorobimane (MCB), which forms an adduct with glutathione in a reaction catalyzed by glutathione-S-Transferase (GST). The unbound MCB is almost nonfluorescent, whereas it emits a fluorescent blue light (Ex/Em = 380nm/461nm) when bound to reduced or oxidized glutathione. Thus, the amount of glutathione can be easily detected using a fluorometer or a 96-well fluorometric plate reader.
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Notes
This product is manufactured by BioVision, an Abcam company and was previously called K251 Glutathione Fluorometric Assay Kit. K251-100 is the same size as the 100 test size of ab65322.
Glutathione (GSH) is a tripeptide that contains L-cysteine, L-glutamic acid and glycine. It is the smallest intracellular protein thiol molecule in the cells, which prevents cell damage caused by reactive oxygen species such as free radicals and peroxides. Glutathione exists in reduced (GSH) and oxidized (GSSG) states. Reduced glutathione (GSH) is a major tissue antioxidant that provides reducing equivalents for the glutathione peroxidase (GPx) catalyzed reduction of lipid hydroperoxides to their corresponding alcohols and hydrogen peroxide to water. In the GPx catalyzed reaction, the formation of a disulfide bond between two GSH molecules generates oxidized glutathione (GSSG). The enzyme glutathione reductase (GR) recycles GSSG to GSH with the simultaneous oxidation of β-nicotinamide adenine dinucleotide phosphate (β-NADPH2). In healthy cells, more than 90% of the total glutathione pool is in the reduced form (GSH). When cells are exposed to increased levels of oxidative stress, GSSG accumulates and the ratio of GSSG to GSH increases. An increased ratio of GSSG-to-GSH is an indication of oxidative stress. The monitoring of reduced and oxidized GSH in biological samples is essential for evaluating the redox and detoxification status of the cells and tissues against oxidative and free radicals mediated cell injury.
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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 GSH Cell Lysis Buffer 1 x 25ml GSH Standard 1 vial GST Enzyme Mix 1 x 200µl MCB Probe 1 x 200µl -
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. Glutathione is the principal intracellular low-molecular-weight thiol that plays a critical role in the cellular defense against oxidative and nitrosative stress in mammalian cells. Diminished glutathione levels have been observed in the early stages of apoptosis. -
Alternative names
- GSH
- GSSG
Associated products
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Related Products
Images
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Glutathione pool measured in THP-1 macrophages: uninfected cells; WT: infected with M.tuberculosis wild type; KO: infected with M.tuberculosis OppD knock-out; COM: infected with M.tuberculosis OppD knock-out complemented with OppDA gene. 106 cells were infected and lysed by treating them with 100µl of ice cold lysis buffer. Cell lysate was diluted and mixed as described in the kit protocol. After 30 min incubation at 37C, fluorescence was measured at Ex=380nm/ Em=460nm. Results represent the means of ± S.D. of three determinations.
Image obtained from Dasgupta A. et al; PLoS One; 2010 Aug 17; 5(8): e12225. -
Glutathione assays were performed using various amounts of Glutathione as indicated. Results were analyzed according to the kit instructions.
Datasheets and documents
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SDS download
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Datasheet download
References (34)
ab65322 has been referenced in 34 publications.
- Wan Y et al. Long noncoding RNA ADAMTS9-AS1 represses ferroptosis of endometrial stromal cells by regulating the miR-6516-5p/GPX4 axis in endometriosis. Sci Rep 12:2618 (2022). PubMed: 35173188
- Tan H et al. Glabridin, a bioactive component of licorice, ameliorates diabetic nephropathy by regulating ferroptosis and the VEGF/Akt/ERK pathways. Mol Med 28:58 (2022). PubMed: 35596156
- Hossain MM et al. Deltamethrin-Evoked ER Stress Promotes Neuroinflammation in the Adult Mouse Hippocampus. Cells 11:N/A (2022). PubMed: 35741090
- Abu Helal R et al. Aortic Fibrosis in Insulin-Sensitive Mice with Endothelial Cell-Specific Deletion of Ceacam1 Gene. Int J Mol Sci 23:N/A (2022). PubMed: 35457157
- Shil A et al. Investigating the Opposing Effect of Two Different Green Tea Supplements on Oxidative Stress, Mitochondrial Function and Cell Viability in HepG2 Cells. J Diet Suppl 19:459-482 (2022). PubMed: 33729080