Gsh gssg glo luminescent assay

Manufactured by Promega

Sourced in United Kingdom

The GSH/GSSG-Glo luminescent assay is a lab equipment product from Promega. It is designed to measure the levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) in biological samples.

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Lab products found in correlation

Colorimetric assay, by Trinity Biotech (1 mentions) White walled 96 well plates, by Promega (1 mentions) Explorer luminometer, by Promega (1 mentions)

Spelling variants of this product

GSH/GSSG-Glo (20 citations) GSH-Glo assay (20 citations) Luminescent assays (2 citations)

2 protocols using gsh gssg glo luminescent assay

Intracellular Lactate and ATP Quantification

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Lactate concentration was determined by colorimetric assay (Trinity Biotech, Bray, Ireland) and compared to a standard curve of known lactate concentrations. The lactate present as a result of efflux from the cell was determined by measuring cell free samples and deducting the amount of lactate originally in the media (derived from the FCS). Intracellular lactate was evaluated using the same assay; cells were washed in PBS prior to lysis in Tris/NaCl lysis buffer (20mM Tris, 150mM NaCl, 1% Triton-X100, pH 7.6).
The ATP concentration in cell lysates was determined by a luminescent assay (ENLITEN ATP system, Promega, Southampton, UK) and comparison made to the cellular ATP concentrations in lysates derived from untreated normoxic controls.
GSH and GSSG concentrations were determined in the same lysates by GSH/GSSG-Glo luminescent assay (Promega, Southampton, UK).

Bola B.M., Chadwick A.L., Michopoulos F., Blount K.G., Telfer B.A., Williams K.J., Smith P.D., Critchlow S.E, & Stratford I.J. (2014). Inhibition of Monocarboxylate transporter-1 (MCT1) by AZD3965 enhances radiosensitivity by reducing lactate transport. Molecular cancer therapeutics, 13(12), 2805-2816.

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Metformin Impact on Cellular GSH/GSSG

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Cells were seeded in cRPMI in white-walled 96-well plates (Promega, Madison, WI, USA) at a density of 15,000 cells/well. At 24 h post seeding, medium was removed to waste, and cells were treated with 100 µL of metformin (10 mM) or vehicle control (H₂O) diluted in cRPMI and incubated at 37°C in 5% CO₂/95% humidified air. After 24 h treatment, media was removed to waste and GSH/GSSG levels measured using the GSH/GSSG-Glo™ luminescent assay (Promega), according to the manufacturer’s instructions. Luminescence was measured at 1 s integration time using the Explorer Luminometer (Promega). An identical plate of cells was set up and treated as with the experimental plate and was used for normalisation of assay results by crystal violet assay.

Buckley C.E., O’Brien R.M., Nugent T.S., Donlon N.E., O’Connell F., Reynolds J.V., Hafeez A., O’Ríordáin D.S., Hannon R.A., Neary P., Kalbassi R., Mehigan B.J., McCormick P.H., Dunne C., Kelly M.E., Larkin J.O., O’Sullivan J, & Lynam-Lennon N. (2023). Metformin is a metabolic modulator and radiosensitiser in rectal cancer. Frontiers in Oncology, 13, 1216911.

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