After the successful launch of our
SensoLyte®
520 Thiol Quantitation Assay Kit (patent pending), AnaSpec received
several inquiries for a sensitive glutathione (GSH) assay. In response, AnaSpec
is pleased to release the SensoLyte®
520 Total GSH Assay Kit (patent pending).
The SensoLyte®
520 Total GSH Assay Kit provides a convenient and highly sensitive fluorescent
assay for the measurement of total GSH (GSH+GSSG). It employs a novel fluorogenic
reagent, which upon reaction with reduced GSH releases fluorescence. Fluorescence
is monitored at Ex/Em= 490/520 nm. As well, oxidized GSH can be converted to
the reduced GSH by glutathione reductase and total GSH level quantified. The
assay has a linear range of 0.2-100 mM of glutathione. The kit
also provides sulfosalicylic acid for deproteination of biological samples.
A colorimetric GSH kit, the SensoLyte®
Total GSH Assay Kit is also available. This kit contains Ellman’s Reagent
[5, 5’-dithiobis (2-nitrobenzoic acid)], which upon reaction with reduced GSH
releases a colored product, monitored at 415 nm using a microplate reader. The
intensity of the color produced is proportional to GSH concentration.
Reduced Glutathione (GSH) is the main thiol (sulfhydryl)
compound in animal, plant and protozoa. GSH plays a crucial role in cellular
defense against oxidative stress and functions as a cofactor for a variety of
enzymes such as glutathione transferase and glutathione peroxidase. GSH is also
involved in the maintenance of redox status, amino acid transport, and mitochondrial
respiration.1,2 Total GSH includes the reduced and oxidized form
(GSSG). Upon oxidation, GSH is converted to GSSG, with the
ratio of GSH/GSSG decreasing due to GSSG accumulation. The concentrations
of GSH and GSSG and their molar ratio are indicators of the cell’s functionality
and oxidative stress level.3, 4
| |
SensoLyte®
520 Total GSH Assay |
SensoLyte®
Total GSH Assay |
| Read-out |
Fluorimetric |
Colorimetric |
| Wavelengths |
Ex/Em= 490/520 nm |
Abs=415 nm |
| Hydrolysis
Product |
A green fluorophore |
2-Nitro-5-thiobenzoic acid (TNB) |
| Dynamic
Linear Range |
0.2-100 mM |
1.5-100 mM |
| Kit
size
(96-well plate) |
100 tests |
500 tests |
| Standard
curve |
|
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Related Products
References
1.
Dickinson, DA. et al. Ann. N. Y. Acad. Sci. 937, 488 (2002).
2. Townsend,
DM. et al. Biomed. Oharmacother. 57, 145 (2003).
3. Ballatori,
N. et al. Biol. Chem. 390, 191 (2009).
4. Rahman,
I. et al. Antioxid. Redox Signal. 7, 42 (2005).
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