GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) ab138881 is used to quantitate reduced and oxidized glutathione (GSH/GSSG).
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- Molecular medicine (Cambridge, Mass.) 30:242024Porous Se@SiO nanospheres alleviate diabetic retinopathy by inhibiting excess lipid peroxidation and inflammation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTian Niu,Xin Shi,Xijian Liu,Haiyan Wang,Kun Liu,Yupeng XuPubMed 38321393
- Experimental and therapeutic medicine 27:262023Breviscapine attenuates lead‑induced myocardial injury by activating the Nrf2 signaling pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDexuan Li,Zhengliang Xu,Yashan Li,Yanmei Huang,Jiali Yin,Hongjuan Li,Beiji ZhangPubMed 38125346
- The Journal of clinical investigation 133:2023Aryl hydrocarbon receptor sulfenylation promotes glycogenolysis and rescues cancer chemoresistance.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNannan Zhou,Jie Chen,Zheng Ling,Chaoqi Zhang,Yabo Zhou,Dianheng Wang,Li Zhou,Zhenfeng Wang,Nan Sun,Xin Wang,Huafeng Zhang,Ke Tang,Jingwei Ma,Jiadi Lv,Bo HuangPubMed 38099490
- Scientific reports 13:201452023Deferoxamine attenuates visual impairment in retinal ischemia‒reperfusion via inhibiting ferroptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXiaoxuan Wang,Mingran Li,Ke Diao,Yan Wang,Hong Chen,Ziqi Zhao,Yuan Li,Xin Jia,Hao Wang,Fangyuan Zheng,Zihan Xia,Longhui Han,Minglian ZhangPubMed 37978208
- Redox biology 68:1029582023A role for the cystathionine-β-synthase /HS axis in astrocyte dysfunction in the aging brain.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAnindya Dey,Pijush Kanti Pramanik,Shailendra Kumar Dhar Dwivedi,Fiifi Neizer-Ashun,Tamas Kiss,Abhrajit Ganguly,Heather Rice,Priyabrata Mukherjee,Chao Xu,Mohiuddin Ahmad,Anna Csiszar,Resham BhattacharyaPubMed 37948927
- International journal of molecular sciences 24:2023Age-Related Shift in Cardiac and Metabolic Phenotyping Linked to Inflammatory Cytokines and Antioxidant Status in Mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRyeonshi Kang,Charlotte Laborde,Lesia Savchenko,Audrey Swiader,Nathalie Pizzinat,Dimitri Marsal,Yannis Sainte-Marie,Frederic Boal,Helene Tronchere,Jerome Roncalli,Oksana KunduzovaPubMed 37958823
- International journal of molecular sciences 24:2023Effect of Antioxidant Supplementation on NET Formation Induced by LPS In Vitro; the Roles of Vitamins E and C, Glutathione, and N-acetyl Cysteine.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGermán Muñoz-Sánchez,Lucila A Godínez-Méndez,Mary Fafutis-Morris,Vidal Delgado-RizoPubMed 37685966
- Methods in molecular biology (Clifton, N.J.) 2712:233-2512023Detection of Ferroptosis in Models of Brain Diseases.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDanmin Shen,Fei Yang,Qian LiPubMed 37578711
- Cancers 15:2023Late Effects of Chronic Low Dose Rate Total Body Irradiation on the Heart Proteome of ApoE Mice Resemble Premature Cardiac Ageing.Applications:Unspecified applicationReactive species:Unspecified reactive speciesOmid Azimzadeh,Juliane Merl-Pham,Vikram Subramanian,Kateryna Oleksenko,Franziska Krumm,Mariateresa Mancuso,Emanuela Pasquali,Ignacia B Tanaka,Satoshi Tanaka,Michael J Atkinson,Soile Tapio,Simone MoertlPubMed 37444528
- Discover. Oncology 14:422023RUNX1-IT1 favors breast cancer carcinogenesis through regulation of IGF2BP1/GPX4 axis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShengting Wang,Yufang Wang,Qian Li,Kaixuan Zeng,Xiaoming Li,Xinghua FengPubMed 37036576
Key facts
- Detection method
- Fluorescent
- Sample types
- Urine, Plasma, Tissue Extracts, Serum, Cell Lysate
- Assay type
- Quantitative
- Assay time
- 30m
- Sensitivity
- = 10 nM
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GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) ab138881 is used to quantitate reduced and oxidized glutathione (GSH/GSSG).
Key facts
- Detection method
- Fluorescent
- Sample types
- Urine, Plasma, Tissue Extracts, Serum, Cell Lysate
- Assay type
- Quantitative
- Assay time
- 30m
- Assay Platform
- Microplate reader
- Sensitivity
- = 10 nM
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
How the assay works
The GSH/GSSG assay protocol uses a proprietary non-fluorescent dye that becomes strongly fluorescent upon reacting with GSH. With a one-step fluorimetric method, the assay can detect as little as 1 picomole of GSH or GSSG in a 100 µL assay volume.
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).
Glutathione reductase (GR) recycles GSSG to GSH with the simultaneous oxidation of ß-nicotinamide adenine dinucleotide phosphate (ß-NADPH2).
In healthy cells, >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 assay can be performed in a convenient 96-well or 384-well microtiter-plate format and readily adapted to automation without a separation step. Its signal can be easily read by a fluorescence microplate reader at Ex/Em = 490/520 nm.
GSH/GSSG assay protocol summary:
- add samples (deproteinized) and standards to wells
- for GSH assay add Thiol Green in assay buffer, or for total glutathione (GSH + GSSG) assay also add GSSG probe
- incubate for 10 - 60 min at room temp
- analyze with microplate reader
GSSG levels can be calculated by subtracting GSH from total glutathione levels.
NOTE: For measuring GSH Standard only, there is enough reagent provided to perform 200 tests.
Alternative assay kits
As an alternative to this kit, we recommend GSH/GSSG Ratio Detection Assay Kit II (Fluorometric - Green) (GSH/GSSG Ratio Detection Assay Kit II (Fluorometric - Green) ab205811) that is a reformulated version of ab138881 with similar performance. It uses a water-soluble probe, which is more stable, than the DMSO-soluble dye used in ab138881.
How other researchers are using
GSH/GSSG Ratio Detection Assay Kit has been used in a variety of sample type including:
- Human retinal microvascular endothelial cells 1
- Rat cardiomyocyte-derived H9C2 cells 2
- Murine B16 melanoma, human MCF-7 breast cancer, SW1990 pancreatic carcinoma cell lines 3
References:
1 - Niu T et al. 2024; 2 - Li D et al. 2024; 3 - Zhou N at al. 2023.
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9 product images
Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
Total GSH measured in cell lysates showing quantity (umol) per 1 mln cells.
Samples with the concentration of 1e7-1e8 cells/mL were used. Samples were diluted 10-1000 fold.
Biesemann, Nadine et al., Scientific reports: vol. 8,1 9408., Fig 4, doi:10.1038/s41598-018-27614-8Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
Measurement of reduced (GSH) and oxidized (GSSG) forms of glutathione in myotubes treated with 150 μM TBHP for 1 h (n = 6).
Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
GSH in reduced state measured in cell lysates showing quantity (umol) per 1 mln cells.
Samples with the concentration of 1e7-1e8 cells/mL were used. Samples were diluted 10-1000 fold.
Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
GSH in reduced state measured in tissue lysates showing quantity (umol) per miligram of extracted protein of tested sample.
Protein concentration for samples varied from 6 mg/mL to 16 mg/mL. Samples were diluted 10-100 fold.
Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
Total GSH measured in tissue lysates showing quantity (umol) per miligram of extracted protein of tested sample.
Protein concentration for samples varied from 6 mg/mL to 16 mg/mL. Samples were diluted 10-100 fold.
Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
GSH in reduced state measured in biological fluids showing concentration (uM) in tested samples. Human samples were diluted 10 fold. Rat sample was diluted 10-1000 fold.
Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
Total GSH measured in biological fluids showing concentration (uM) in tested samples. Samples were diluted 10-100 fold.
Biochemical assay - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
Niu et al used Lipid Peroxidation (MDA) Assay Kit Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) ab118970, GSH/ GSSG Assay Kit ab138881, and Glutathione Peroxidase Assay Kit Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] ab125066 to investigate the use of porous Se@SiO2 nanospheres to achieve controlled release of selenium, a scavenger of intracellular free radicals, in mouse eyes.
Male diabetic db/db and control db/m mice were injected in the eyes with porous Se@SiO2 nanospheres, and porous SiO2 nanospheres (NPs) without Se.
Porous Se@SiO2 nanospheres inhibit diabetes-induced retinal lipid peroxidation and inflammation. Levels of MDA in retinal homogenates (n = 6). Expression levels of GPX4 in retinas were measured using western blotting; β‐actin was used as a loading control (left panel). Band densities were assessed using the ImageJ software, and GPX4 expression levels are represented as their ratios to β‐actin (right panel) (n = 3). Levels of GSH, GSSG, and the ratio of GSH to GSSG in retinal homogenates (n = 6). Data are represented as the mean ± SD. **p < 0.01, ***p < 0.001; ns, nonsignificant.
Retinal samples were harvested, washed, and lysed according to the manufacturer’s instructions. Protein concentrations were determined using a bicinchoninic acid kit. MDA and glutathione concentrations were determined using a lipid peroxidation MDA assay kit, reduced glutathione (GSH) / oxidized glutathione (GSSG) Ratio Detection Assay kit (Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) ab118970, ab138881; Abcam, MA, USA), and a microplate reader. The relative concentrations of MDA and glutathione were calculated by normalizing the measured concentrations to that of the total protein.
Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881)
GSH and Total (GSH+GSSG) dose responses were measured with ab138881. Blue line: in the presence of GSH only; Red line: in the presence of 1:1 GSH/GSSG.
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