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Glutathione Assay Kit (Fluorometric) (ab65322) provides a simple in vitro assay for detection of total glutathione changes during cellular response to toxicity, apoptosis and other conditions.


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Functional Studies - Glutathione Assay Kit (Fluorometric) (AB65322), expandable thumbnail
  • Functional Studies - Glutathione Assay Kit (Fluorometric) (AB65322), expandable thumbnail

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Key facts

Detection method

Fluorescent

Sample types

Urine, Plasma, Tissue Extracts, Cell culture media, Serum, Other biological fluids, Cell Lysate

Assay type

Quantitative

Reactive species

Mammals

Assay time

2h

Target data

What's included?

100 Test
Components
GSH Cell Lysis Buffer
1 x 25 mL
GSH Standard
1 x 1 Vial
GST Enzyme Mix
1 x 200 µL
MCB Probe
1 x 200 µL

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Glutathione Assay Kit (Fluorometric) (ab65322) provides a simple in vitro assay for detection of total glutathione changes during cellular response to toxicity, apoptosis and other conditions.

Key facts

Detection method

Fluorescent

Sample types

Urine, Plasma, Tissue Extracts, Cell culture media, Serum, Other biological fluids, Cell Lysate

Assay type

Quantitative

Reactive species

Mammals

Assay time

2h

Assay Platform

Microplate reader

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

Glutathione Assay Kit (Fluorometric) (ab65322) provides a simple in vitro assay for detection of total glutathione changes during cellular response to toxicity, apoptosis and other conditions. The assay uses the dye monochlorobimane (MCB), which forms an adduct with glutathione in a reaction catalyzed by glutathione-S-Transferase (GST). The unbound MCB is almost nonfluorescent, whereas it emits a fluorescent blue light (Ex/Em = 380nm/461nm) when bound to reduced or oxidized glutathione. Thus, the amount of glutathione can be easily detected using a fluorometer or a 96-well fluorometric plate reader.

Visit our FAQs page for tips and troubleshooting.

This product is manufactured by BioVision, an Abcam company and was previously called K251 Glutathione Fluorometric Assay Kit. K251-100 is the same size as the 100 test size of ab65322.

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). The enzyme glutathione reductase (GR) recycles GSSG to GSH with the simultaneous oxidation of β-nicotinamide adenine dinucleotide phosphate (β-NADPH2). In healthy cells, more than 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 monitoring of reduced and oxidized GSH in biological samples is essential for evaluating the redox and detoxification status of the cells and tissues against oxidative and free radicals mediated cell injury.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Glutathione often referred to as GSH is a small tripeptide with a molecular mass of approximately 307.33 g/mol. It is composed of amino acids glutamine cysteine and glycine. Glutathione is found in almost all cells with high concentrations especially in the liver. As a powerful antioxidant it functions by reducing reactive oxygen species (ROS) and maintaining the redox status of cells. Glutathione exists in two forms: reduced (GSH) and oxidized (GSSG). The balance between these forms known as the GSH/GSSG ratio is critical for cellular health and is commonly assessed using a glutathione assay kit.

Biological function summary

Glutathione plays an important role in detoxification processes protecting cells from damage caused by free radicals peroxides and heavy metals. It acts as a substrate for various enzymes like glutathione peroxidase and glutathione S-transferase involved in neutralizing oxidative damage. Glutathione doesn't typically form a complex but it does participate in disulfide bond exchange reactions which are critical in maintaining cellular protein functions and structures.

Pathways

Glutathione is integral to cellular antioxidant defense and phase II detoxification pathways. It closely interacts with proteins like glutathione peroxidase in the reduction of hydrogen peroxide to water. Also glutathione's role in the pentose phosphate pathway supports the regeneration of NADPH which is essential for maintaining the reduced form of glutathione. This interplay is important for protecting cells against oxidative stress and ensuring cellular energy is balanced.

Associated diseases and disorders

Glutathione has significant implications in conditions such as Parkinson's disease and liver cirrhosis. In Parkinson's disease researchers observe lower levels of glutathione in affected brain regions suggesting an important role in neurodegeneration. Glutathione levels also impact liver health with deficiencies leading to ineffective detoxification and the potential for liver damage or cirrhosis. In these contexts glutathione interacts with proteins involved in redox balance and neurotransmitter regulation emphasizing its broad significance in maintaining human health.

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2 product images

  • Functional Studies - Glutathione Assay Kit (Fluorometric) (ab65322), expandable thumbnail
    Dasgupta A et al., PLoS One, 5, , 2010 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Glutathione Assay Kit (Fluorometric) (ab65322)

    Glutathione pool measured in THP-1 macrophages:
    uninfected cells;


    WT: infected with M.tuberculosis wild type;


    KO: infected with M.tuberculosis OppD knock-out;


    COM: infected with M.tuberculosis OppD knock-out complemented with OppDA gene.


    106 cells were infected and lysed by treating them with 100μl of ice cold lysis buffer. Cell lysate was diluted and mixed as described in the kit protocol. After 30 min incubation at 37C, fluorescence was measured at Ex=380nm/ Em=460nm. Results represent the means of ± S.D. of three determinations.

    Image obtained from Dasgupta A. et al; PLoS One; 2010 Aug 17; 5(8): e12225.

  • Functional Studies - Glutathione Assay Kit (Fluorometric) (ab65322), expandable thumbnail

    Functional Studies - Glutathione Assay Kit (Fluorometric) (ab65322)

    Glutathione assays were performed using various amounts of Glutathione as indicated. Results were analyzed according to the kit instructions.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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