Glutathione Assay Kit

Cayman’s GSH assay kit utilizes a carefully optimized enzymatic recycling method, using glutathione reductase for the quantification of GSH.^1,2,3 The sulfhydryl group of GSH reacts with DTNB (5,5'-dithio-bis-2-nitrobenzoic acid, Ellman’s reagent) and produces a yellow colored 5-thio-2-nitrobenzoic acid (TNB). The mixed disulfide, GSTNB (between GSH and TNB) that is concomitantly produced, is reduced by glutathione reductase to recycle the GSH and produce more TNB. The rate of TNB production is directly proportional to this recycling reaction which in turn is directly proportional to the concentration of GSH in the sample. Measurement of the absorbance of TNB at 405 or 412 nm provides an accurate estimation of GSH in the sample. GSH is easily oxidized to the disulfide dimer GSSG. Because of the use of glutathione reductase in the Cayman GSH assay kit, both GSH and GSSG are measured and the assay reflects total glutathione. The kit can also be used to measure only GSSG by following an alternative protocol. GSH measurement can be done in plasma, tissue samples, and cultured cells using this kit. Nearly all samples require deproteination before assay.

Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.

WARNING This product is not for human or veterinary use.

1. Baker, M.A., Cerniglia, G.J., and Zaman, A. Microtiter plate assay for the measurement of glutathione and glutathione disulfide in large numbers of biological samples. Anal. Biochem. 190, 360-365 (1990).

2. Eyer, P., and Podhradsky, D. Evaluation of the micromethod for determination of glutathione using enzymatic cycling and Ellman’s reagent. Anal. Biochem. 153, 57-66 (1986).

3. Tietze, F. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues. Anal. Biochem. 27, 502-522 (1969).

Camilliere, M., Verde, M.R., Rabadi, S.M., et al. Maternal undernourishment impairs murine placental development during pregnancy. BMC Pregnancy Childbirth 25(1), 889 (2025).

Wu, K., Yan, M., Liu, T., et al. Creatine kinase B suppresses ferroptosis by phosphorylating GPX4 through a moonlighting function. Nat. Cell. Biol. 25(5), 714-725 (2023).

Li, Z., Ferguson, L., Deol, K.K., et al. Ribosome stalling during selenoprotein translation exposes a ferroptosis vulnerability. Nat. Chem. Biol. (2022).

Kalamkar, S., Acharya, J., Madathil, A.K., et al. Randomized clinical trial of how long-term glutathione supplementation offers protection from oxidative damage and improves HbA1c in elderly type 2 diabetic patients. Antioxidants (Basel) 11(5), 1026 (2022).

Armenta, D.A., Laqtom, N.N., Alchemy, G., et al. Ferroptosis inhibition by lysosome-dependent catabolism of extracellular protein. Cell Chem. Biol. 29(11), 1588-1600 (2022).

Conlon, M., Poltorack, C.D., Forcina, G.C., et al. A compendium of kinetic modulatory profiles identifies ferroptosis regulators. Nat. Chem. Biol. (2021).

Yang, Y., Luo, M., Zhang, K., et al. Nedd4 ubiquitylates VDAC2/3 to suppress erastin-induced ferroptosis in melanoma. Nat. Commun. 11(1), 433 (2020).

Selvakumar, G.P., Ahmed, M.E., Thangavel, R., et al. A role for glia maturation factor dependent activation of mast cells and microglia in MPTP induced dopamine loss and behavioural deficits in mice. Brain Behav. Immun. 87, 429-443 (2020).

Bersuker, K., Hendricks, J., Li, Z., et al. The CoQ oxidoreductase FSP1 acts parallel to GPX4 to inhibit ferroptosis. Nature 575(7784), 688-692 (2019).

Cao, J.Y., Poddar, A., Magtanong, L., et al. A genome-wide haploid genetic screen identifies regulators of glutathione abundance and ferroptosis sensitivity. Cell Rep. 26(6), 1544-1556 (2019).

Magtanong, L., Ko, P.-J., To, M., et al. Exogenous monounsaturated fatty acids promote a ferroptosis-resistant cell state. Cell Chem. Biol. 26(3), 420-432 (2019).

Bian, Y., Kim, K., An, G.-J., et al. Dapsone hydroxylamine, an active metabolite of dapsone, can promote the procoagulant activity of red blood cells and thrombosis. Toxicol. Sci. 172(2), 435-444 (2019).

Venkatesan, T., Choi, Y.-W., and Kim, Y.-K. Effect of an extraction solvent on the antioxidant quality of Pinus densiflora needle extract. J. Pharm. Anal. 9(3), 193-200 (2019).

Schulte, M.L., Fu, A., Zhao, P., et al. Pharmacological blockade of ASCT2-dependent glutamine transport leads to antitumor efficacy in preclinical models. Nat. Med. 24(2), 194-202 (2018).

Wu, M., Yang, Y., Wang, M.Z., F., et al. Exogenous pancreatic kallikrein improves diabetic cardiomyopathy in streptozotocin-induced diabetes. Front. Pharmcacol. 9:855, (2018).

Kim, Y.S., Chung, Y.H., Seo, D.S., et al. Twenty-eight-day repeated inhalation toxicity study of aluminum oxide nanoparticles in male Sprague-Dawley rats. Toxicol. Res. 34(4), 343-354 (2018).

Wang, M.-Y., Srinivasan, M., Dasari, D., et al. Antioxidant activity of Yichun Blue Honeysuckle (YBHS) berry counteracts CCl4-Induced toxicity in liver injury model of mice. Antioxidants 6(3), 50 (2017).

Alarifi, S., Ali, H., Alkahtani, S., et al. Regulation of apoptosis through bcl-2/bax proteins expression and DNA damage by nano-sized gadolinium oxide. Int. J. Nanomedicine 12, 4541-4551 (2017).

Velviranli, M., Okudan, N., Revan, S., et al. Repeated supramaximal exercise-induced oxidative stress: Effect of β-alanine plus creatine supplementation. Asian J. Sports Med. 7(1), e26843 (2016).

Salvi, A., Patki, G., Khan, E., et al. Relationship between advanced glycation end products and increased lipid peroxidation in semen of diabetic men. Oxid. Med. Cell. Longev. 5059043 (2016).

Davies, T.G., Wixted, W.E., Coyle, J.E., et al. Monoacidic inhibitors of the Kelch-like ECH-associated protein 1: Nuclear factor erythroid 2-related factor 2 (KEAP1:NRF2) Protein-protein interaction with high cell potency identified by fragment-based discovery. J. Med. Chem. 59(8), 3991-4006 (2016).

Liu, X., Zhang, S., Whitowrth, R.J., et al. Unbalanced activation of glutathione metabolic pathways suggests potential involvement in plant defense against the gall midge Mayetiola destructor in wheat. Sci. Rep. 5:8092, (2015).

Faisal, M., Saquib, Q., Alatar, A.A., et al. Phytotoxic hazards of NiO-nanoparticles in tomato: A study on mechanism of cell death. J. Hazard. Mater. 250-251, 318-332 (2013).

Liu, J.Q., Lee, T.F., Bigam, D.L., et al. Effects of post-resuscitation treatment with N-acetylcysteine on cardiac recovery in hypoxic newborn piglets. PLoS One 5(12), e15322 (2010).

Liu, J.-Q., Lee, T.-F., Bigam, D.L., et al. Effects of post-resuscitation treatment with N-acetylcysteine on cardiac recovery in hypoxic newborn piglets. PLoS One 5(12), e15322 (2010).