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

1-10 of 16 Abreviews or Q&A

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

Answer

Thank you for your inquiry.

I am happy to confirm that ab102530 (Glutathione Peroxidase Assay Kit) is suitable for all mammalian samples, including rat samples.

I wish your customer good luck with their experiments.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of a new lot in addition to the positive control with the order number xxxxxxxxx.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

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Answer

I have recieved a reply from the lab who confirm that there was a problem with the positive control from this lot. Unfortunately the positive control from this lot lost activity for unforeseen reasons
I am sorry this product did not perform as stated on the datasheet and for the inconvenience that this has caused.
As a compensation I am delighted to issue you with a free of charge replacement of the positive control from a different lot.
The new order number is xxxxxxxx.
Please check that the below shipping address is correct:


To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

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Answer

Thank you for contacting us.
I am currently in contact with the lab who are rechecking the positive control from this lot.
In the meantime, they would like to know what difference you got in the absorbance from the vector alone and the GPx expressing vector? If there is an insignificant difference, it might just be experimental artifact and have nothing to do real values.
Another possibility is that the GPx from the vector is interfering with the readings from the endogenous GPx and hence you see relatively higher reading from the empty vector
I hope this information is helpful to you and I look forward to your response.
Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for your inquiry.

I am sorry to confirm that we do not offer the positive control separately for purchase.

I am sorry to not have a positive answer on this occasion and wish you a good weekend.

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Answer

Thank you for contacting us.

Are you planning to use antibodies for detection of these enzyme. Which application you would be interested in e.g. Immunohistochemistry or western blotting. If you are planning to use antibodies then please use are website there are range of antibodies available as per your requirements.

The following kits I can recommend for assaying the activity of these enzyme;

https://www.abcam.com/glutathione-reductase-gr-assay-kit-ab83461.html

https://www.abcam.com/glutathione-peroxidase-assay-kit-ab102530.html

https://www.abcam.com/superoxide-dismutase-activity-colorimetric-assay-kit-ab65354.html

https://www.abcam.com/catalase-assay-kit-ab83464.html

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

Could you please clarify; do you want to detect these markers in your tissue samples or you just want to assay the activity of these enzymes?

I will be looking forward to hearing from you soon.

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Answer

Thank you for contacting us.

Abcam takes product quality seriously and we recently undertook an in-depth study examining storage conditions and antibody performance. After over 13,000 individual ELISA experiments, we have determined that storing our antibodies at various temperatures of up to 45°C for 1 week does not impact their activity. I would be happy to share a summary of our findings with you; please find the results attached.

As a precautionary measure only, we ship our antibodies in packaging with ice packs to provide extra temperature stability in transit. The ice pack may be thawed or even at room temperature when you receive it; please be assured that this is normal and that your product is safe to use. Once you have received the vial, please follow the long-term storage instructions on the datasheet.

Our Abpromise to you is that, should our product not work as stated on the datasheet, we will resolve the issue to your satisfaction, either with helpful advice to optimize your experiment or a replacement or refund of the product. Please contact our knowledgeable Scientific Support staff should you experience any issues while using our products. Contact information can be found at:
https://www.abcam.com/index.html?pageconfig=contactus

I hope this information helps, but please don't hesitate to contact me for additional information or assistance.

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Answer

Thank you for contacting us.

Please do not get confused with cytosol and mitochondrial SODs for plasma; the information I had provided was for general cells samples. I had missed the fact you are going to use plasma and plasma does not contain cells.

You would not need ab65354.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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1-10 of 16 Abreviews or Q&A

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

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