Overview

  • Product name
    Glutathione Peroxidase Assay Kit (Colorimetric)
    See all Glutathione Peroxidase kits
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Platelets, Other biological fluids, Tissue Extracts
  • Assay type
    Enzyme activity
  • Sensitivity
    0.5 mU/ml
  • Assay time
    0h 40m
  • Species reactivity
    Reacts with: Mammals, Other species
  • Product overview

    Glutathione Peroxidase Assay Kit (Colorimetric) ab102530 can be used to quantitate the activity of all of the glutathione dependent peroxidases in plasma, erythrocyte lysates, tissue homogenates, and cell lysates.


    In the glutathione peroxidase assay protocol, glutathione peroxidase (GPx) oxidizes GSH to produce GSSG as part of the reaction in which it reduces cumene hydroperoxide. Glutathione reductase (GR) then reduces the GSSG to produce GSH, and in the same reaction consumes NADPH. The decrease of NADPH (measured at OD=340 nm) is proportional to GPx activity. 


    The assay has a detection sensitivity of ~ 0.5 mU/ml of GPx in samples.

    Glutathione peroxidase assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 15 min at room temp to deplete all GSSG in sample
    - add cumene hydroperoxide
    - analyze with microplate reader immediately, and after at least 5 min

  • Notes

    The Glutathione Peroxidase (GPx, EC 1.11.1.9) family of enzymes plays and important role in the protection of organisms from oxidative damage. GPx converts reduced glutathione (GSH) to oxidized glutathione (GSSG) while reducing lipid hydroperoxides to their corresponding alcohols or free hydrogen peroxide to water. 

  • Platform
    Microplate reader

Properties

Images

  • Scatter dot plots representing the levels of glutathione peroxidase (GPx) in brain tissue in different mice groups.

  • The effects of PCSE on the activity of GPX in a mouse UC model.
  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted
  • Glutathione Peroxidase Activity measured in biological fluids showing quantity (nmol) per 1 mL after 10 min of incubation. Samples diluted 1-27 fold.

  • Glutathione Peroxidase Activity measured in cell lysates showing quantity (nmol) per 1 mln cells after 10 min of incubation. Undiluted sample contained 2.5e6 cells per well. Samples were diluted 1-9 fold.

  • Tests example obtained using ab102530

Protocols

References

This product has been referenced in:
  • Vande Voorde J  et al. Improving the metabolic fidelity of cancer models with a physiological cell culture medium. Sci Adv 5:eaau7314 (2019). Read more (PubMed: 30613774) »
  • Kaushal JB  et al. Sonic hedgehog protects endometrial hyperplasial cells against oxidative stress via suppressing mitochondrial fission protein dynamin-like GTPase (Drp1). Free Radic Biol Med 129:582-599 (2018). Read more (PubMed: 30347228) »
See all 15 Publications for this product

Customer reviews and Q&As

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Determination of GPx activity in microalgae samples.

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Sample preparation – Microalgae.
1. Harvest the amount of microalgae necessary for each assay (initial recommendation = 15 mg).
2. Resuspend microalgae in 500 μL of cold Assay Buffer.
3. Sonicate the sample for 30 seconds on ice. Repeat three times.
4. Centrifuge 10 minutes at 4ºC at 10,000g using a cold microcentrifuge to remove any insoluble material.
5. Collect supernatant and transfer to a clean tube.
6. Keep on ice.
- It is recommended to perform several dilution of your sample to ensure the reading are within the standard value range. In this case the dilution factor was 1:50.

Laura Baselga

Verified customer

Submitted Aug 31 2018

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

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