SensoLyte® 520 Total GSH Quantitation Assay - Long Wavelength


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


Product

Catalog #

SensoLyte® 520 Total GSH Assay Kit “Fluorimetric”

72154

SensoLyte® Total GSH Assay Kit “Colorimetric”

72153

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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).