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: Mouse, Human, Mammals
  • 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.

Properties

Images

  • Glutathione Detection Kit (Fluorometric) (ab65322)
    Glutathione Detection 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 Detection Kit (Fluorometric) (ab65322)
    Functional Studies - Glutathione Detection Kit (Fluorometric) (ab65322)
    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) »
  • Katerji M  et al. Chemosensitivity of U251 Cells to the Co-treatment of D-Penicillamine and Copper: Possible Implications on Wilson Disease Patients. Front Mol Neurosci 10:10 (2017). Read more (PubMed: 28197071) »

See all 5 Publications for this product

Publishing research using ab65322? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Does this kit measure total glutathione (GSH + GSSG) or just sample GSH concentration?

This kit detects total glutathione.

What plates are recommended for use with this kit?

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.

Hi,

I am using Greiner Thincert Culture Inserts for 6-well plates and A549 cells in my exposure experiments. The goal is to expose the cells at an air/liquid interphase for a certain period and to measure cell viability, oxidative stress mar...

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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 ce...

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Dear technical team, Our customer would like to purchase ab65322 and wants to confirm plasma samples are availble for this kit.   Please check this for our customer.     Best regards,

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...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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