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GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) ab138881 provides an ultrasensitive assay to quantitate glutathione in mammalian samples.

GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) designed for success:
- Simple and fast protocol for the quantitative determination of reduced and oxidized glutathione (GSH/GSSG) in plasma, urine, tissue and cell extracts
- High sensitivity; detection limit can be as low as 1 picomole of GSH or GSSG in a 100 µL assay volume
- Can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step


Images

Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (AB138881), expandable thumbnail
  • Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (AB138881), expandable thumbnail
  • Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (AB138881), expandable thumbnail
  • Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (AB138881), expandable thumbnail
  • Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (AB138881), expandable thumbnail

Publications

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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What's included?

100 Test
Components
Assay Buffer
1 x 25 mL
DMSO
1 x 400 µL
GSH Standard
1 x 62 µg
GSSG Probe
1 x 1 Vial
GSSG Standard
1 x 124 µg
Thiol Green Indicator
1 x 1 Vial

Recommended products

GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) ab138881 provides an ultrasensitive assay to quantitate glutathione in mammalian samples.

GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) designed for success:
- Simple and fast protocol for the quantitative determination of reduced and oxidized glutathione (GSH/GSSG) in plasma, urine, tissue and cell extracts
- High sensitivity; detection limit can be as low as 1 picomole of GSH or GSSG in a 100 µL assay volume
- Can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step

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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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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9 product images

  • Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881), expandable thumbnail

    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.

  • Functional Studies - GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) (ab138881), expandable thumbnail
    Biesemann, Nadine et al., Scientific reports: vol. 8,1 9408., Fig 4, doi:10.1038/s41598-018-27614-8

    Functional 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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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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