Overview

  • Product name

    Glutathione Assay Kit (Fluorometric)
    See all Glutathione kits
  • Detection method

    Fluorescent
  • Sample type

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

    Quantitative
  • Assay time

    2h 00m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    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.


    This product does not measure total glutathione in the sample. It measures the relative level of glutathione between untreated and treated samples.



    Visit our FAQs page for tips and troubleshooting.

  • Notes

    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.

  • Platform

    Microplate reader

Properties

Images

  • 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.

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

Protocols

References

This product has been referenced in:

  • Piao X  et al. 1-Deoxynojirimycin (DNJ) Ameliorates Indomethacin-Induced Gastric Ulcer in Mice by Affecting NF-kappaB Signaling Pathway. Front Pharmacol 9:372 (2018). Read more (PubMed: 29725297) »
  • Naletova I  et al. Cytotoxic phenanthroline derivatives alter metallostasis and redox homeostasis in neuroblastoma cells. Oncotarget 9:36289-36316 (2018). Read more (PubMed: 30555630) »
See all 10 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Question
Answer

This kit detects total glutathione.

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Question
Answer

Black 96-well plates with transparent bottom (flat-bottom wells) can be used for bottom-reading fluorometers or opaque white plates can be used for top-reading fluorometers.

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Answer

Thank you for your enquiry and interest in our products.

MTT assay can be carried out on 96 well plates using either adherent or floating cells. You do not necessary need to remove (harvest) the cells by trypsinization. You can culture the cells in the wells of the 96 well plates, then apply the MTT solution (20ul) to each well. During incubation (37C, 5% CO2) for 1-5 hours MTT will be metabolized. The formazan (MTT metabolic product) can then be dissolved in DMSO and the optical density can be read at 560 nm using a plate reader - without harvesting the cells. There may be other cell viability/cytotoxicyprotovols but this is the most widely used.

Cell viability kits:

We have different cell viability assay kits in our catalogue i.e. ab112120, ab112121, ab112122, ab112123.

https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Blue-ab112120.html

https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Dual-Green-Red-ab112121.html

https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Green-ab112122.html

https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Near-InfraRed-ab112123.html

GSH kits:

GSH can be detected/quantified either by fluorimetric or chromogenic (DTNB - Ellman's reagent) assays and currently we have also some assay kits for detecting this tripeptide, you may wish to take a look at the datasheets for further information.

- ab65322: Glutathione Fluorometric Detection Kit

https://www.abcam.com/Glutathione-Fluorometric-Detection-Kit-ab65322.html

- ab112132:Intracellular GSH Assay Kit

https://www.abcam.com/Intracellular-GSH-Assay-Kit-ab112132.html

Normally, intracellular GSH levels can be expressed per cell numbers or per protein concentration.

I hope this helps and if I can assist further, please do not hesitate to contact me.

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Answer

Thank you for contacting us. The Glutathione Fluorometric Detection Kit (ab65322) can be used with a number of different types of sample: Cells, tissues or liquid samples such as plasma. As long as your sample contains Glutathione which can be detected the kit should be applicable to your customers sample.  A GSH standard is provided with the kit in order to produce the standard curve. I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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