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Glutathione Peroxidase Assay Kit (Colorimetric) (ab102530)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (15)References (52)

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Functional Studies - Glutathione Peroxidase Assay Kit (Colorimetric) (ab102530)
  • Functional Studies - Glutathione Peroxidase Assay Kit (Colorimetric) (ab102530)
  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 40 min
  • Sample type: Cell culture supernatant, Other biological fluids, Plasma, Platelets, Serum, Tissue Extracts, Urine
  • Sensitivity: 0.5 mU/ml

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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

    This product is manufactured by BioVision, an Abcam company and was previously called K762 Glutathione Peroxidase Activity Colorimetric Assay Kit. K762-100 is the same size as the 100 test size of ab102530.

    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

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests 2000 tests
    Cumene Hydroperoxide Yellow 1 vial 20 vials
    Glutathione Brown 1 vial 20 vials
    Glutathione Reductase Green 1 vial 20 vials
    GPx Assay Buffer WM 1 x 50ml 20 x 50ml
    GPx Positive Control Red 1 vial 20 vials
    NADPH Blue 1 vial 20 vials
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Oxidative stress
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Coenzyme and cofactor assay kits
    • Glutathione assay kits
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Antioxidants
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Antioxidant
  • Relevance

    Glutathione Peroxidase (GPx, EC 1.11.1.9) is an enzyme family with peroxidase activity, and plays important role in protecting of organisms from oxidative damage. It converts reduced glutathione (GSH) to oxidized glutathione (GSSG), to reduce lipid hydroperoxides to their corresponding alcohols, or reduce free hydrogen peroxide to water. Several isozymes have been found in different cellular locations and with different substrate specificity. Low levels of GPx have been correlated with free radical related disorders.
  • Alternative names

    • GPX

Images

  • Functional Studies - Glutathione Peroxidase Assay Kit (Colorimetric) (ab102530)
    Functional Studies - Glutathione Peroxidase Assay Kit (Colorimetric) (ab102530)George, Akash K et al., Scientific reports?vol. 8,1 5158., Fig 4, doi:10.1038/s41598-018-23568-z

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

  • Functional Studies - Glutathione Peroxidase Assay Kit (Colorimetric) (ab102530)
    Functional Studies - Glutathione Peroxidase Assay Kit (Colorimetric) (ab102530)Liu, Baohai et al., Frontiers in pharmacology?vol. 9 347., Fig 8, doi:10.3389/fphar.2018.00347
    The effects of PCSE on the activity of GPX in a mouse UC model.
  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
    Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
    Standard curve: mean of duplicates (+/- SD) with background reads subtracted
  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
    Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)

    Glutathione Peroxidase Activity measured in biological fluids showing quantity (nmol) per 1 mL after 10 min of incubation. Samples diluted 1-27 fold.

  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
    Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)

    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.

  • Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
    Functional Studies - Glutathione Peroxidase Assay Kit (ab102530)
    Tests example obtained using ab102530

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (52)

Publishing research using ab102530? Please let us know so that we can cite the reference in this datasheet.

ab102530 has been referenced in 52 publications.

  • Kwon DA  et al. Antioxidant and antifatigue effect of a standardized fraction (HemoHIM) from Angelica gigas, Cnidium officinale, and Paeonia lactiflora. Pharm Biol 59:391-400 (2021). PubMed: 33813987
  • Hayman TJ  et al. STING enhances cell death through regulation of reactive oxygen species and DNA damage. Nat Commun 12:2327 (2021). PubMed: 33875663
  • Chen J  et al. Inhibition of Acyl-CoA Synthetase Long-Chain Family Member 4 Facilitates Neurological Recovery After Stroke by Regulation Ferroptosis. Front Cell Neurosci 15:632354 (2021). PubMed: 33889074
  • Krakowska-Sieprawska A  et al. Promising Green Technology in Obtaining Functional Plant Preparations: Combined Enzyme-Assisted Supercritical Fluid Extraction of Flavonoids Isolation from Medicago Sativa Leaves. Materials (Basel) 14:N/A (2021). PubMed: 34064166
  • El-Nobi G  et al. Synbiotic Effects of Saccharomycescerevisiae, Mannan Oligosaccharides, and ß-Glucan on Innate Immunity, Antioxidant Status, and Disease Resistance of Nile Tilapia, Oreochromis niloticus. Antibiotics (Basel) 10:N/A (2021). PubMed: 34065896
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a question

1-10 of 15 Q&A

Question

Dear All,



Please let us know if the product ab102530-Glutathione Peroxidase can be run on RAT samples.



Early response requested.


Sincere regards,

Read More

Abcam community

Verified customer

Asked on Dec 18 2012

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.

Read More

Abcam Scientific Support

Answered on Dec 18 2012

Question

Thank you for your response regarding my query. However, there is no point in issuing me a positive control as I have already done the experiment, using the other reagents in the kit. I do not have not enough lysate to repeat the assay. If I repeat the whole experiment, with the new positive control, I will effectively be wasting the rest of the kit, which as you will appreciate, is fairly expensive. If I could also be issued the reagents that accompany the positive control, this would be more appropriate.

Read More

Abcam community

Verified customer

Asked on Dec 11 2012

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.

Read More

Abcam Scientific Support

Answered on Dec 11 2012

Question

Inquiry: Hi, I recently tried your GPX assay and found that I got a response from my cells using a density of apprx 2.5 x 10e6. I haven't however got a response for the positive control, using 10ul. I have tried this twice. If you could tell me the concentration of the positive control I'd be grateful/why it is not working. Furthermore, using cells transfected with flag tagged GPX1 and empty vector (containing flag), I get a greater change in absorbance (normalised to protein concentration) with my empty vector than my GPX1 construct. Construct expression have been checked by western blot and the vectors are fully sequenced. Again this has happened twice. There is definitely no mix up in samples as the lysates are from 3 separate experiments. Why would I be getting a stronger reaction with the empty flag vector alone?

Read More

Abcam community

Verified customer

Asked on Dec 07 2012

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.

Read More

Abcam Scientific Support

Answered on Dec 07 2012

Question

Inquiry: Hi, I recently tried your GPX assay and found that I got a response from my cells using a density of apprx 2.5 x 10e6. I haven't however got a response for the positive control, using 10ul. I have tried this twice. If you could tell me the concentration of the positive control I'd be grateful/why it is not working. Furthermore, using cells transfected with flag tagged GPX1 and empty vector (containing flag), I get a greater change in absorbance (normalised to protein concentration) with my empty vector than my GPX1 construct. Construct expression have been checked by western blot and the vectors are fully sequenced. Again this has happened twice. There is definitely no mix up in samples as the lysates are from 3 separate experiments. Why would I be getting a stronger reaction with the empty flag vector alone?

Read More

Abcam community

Verified customer

Asked on Dec 06 2012

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

Read More

Abcam Scientific Support

Answered on Dec 06 2012

Question

Hello,

I would like to ask you about possibility to obtain GPx Positive Control separately. My second question is, what is the catalytic concentration of this control material. Can I have an application sheet?

Thanks a lot

Sincerely yours

Read More

Abcam community

Verified customer

Asked on Nov 16 2012

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.

Read More

Abcam Scientific Support

Answered on Nov 16 2012

Question

Thank you for your reply.

I wish to detect the presence of antioxidant enzymes in the tissue (glutathione peroxidase and reductase, SOD and catalase most important) but also the quantification or activity of each enzyme is important. Essentially I will be comparing tissue from two groups (disease and no disease) and then examining the effect of two drugs:

The total study will be (n = about 6 to 8 tissues per group):

Disease (n=8), No disease (n=8)
Drug A: Group 1 disease (n=8), Group 2 no disease (n=8)
Drug B: Group 3 disease (n=8), Group 4 no disease (n=8)
Drug A + B: Group 5 disease (n=8), Group 6 no disease (n=8)

I appreciate your advice

Read More

Abcam community

Verified customer

Asked on Nov 12 2012

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

Read More

Abcam Scientific Support

Answered on Nov 12 2012

Question

I am inquiring about your total antioxidant capacity assay kit or similar.

I have frozen placental tissue from rabbit experiments and I am wishing to examine markers of oxidative stress.
The most important markers for me are glutathione peroxidase and reductase, superoxide dismutase (SOD) and catalase.

Can you kindly advise me if any of your products are potentially of use and a quote for cost please. I am in Belgium.

Regards,

Read More

Abcam community

Verified customer

Asked on Nov 12 2012

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.

Read More

Abcam Scientific Support

Answered on Nov 12 2012

Question

Hello,
My order# from Abcamencountered problems and is currently experiencing major delays with customs in my country. I want to inquire, will the products in this order still work if they were put for more than 4 days in room temperature?
Here are the list of items in order#:
- Glutathione Peroxidase Assay Kit (ab102530)
- Catalase Assay Kit (ab83464)
- Superoxide Dismutase Colorimetric Assay Kit (ab65354)
- Mitochondria Isolation Kit for Tissue (ab110168)
Please reply ASAP.
Thank you.

Read More

Abcam community

Verified customer

Asked on Jun 22 2012

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.

Read More

Abcam Scientific Support

Answered on Jun 22 2012

Question

Thank you for your email and for the clarification. However, the fractionation kit ab65320 is for use in cells and not plasma, so would not be relevant for our plasma samples. So is this kit still able to measure both Cu/Zn and Mn SOD in our plasma samples? If so what is the protocol for this and would we need 2 ab65354 kits tio measure both SODs in plasma? I hope that is clear now, and looking forward to hearing from you. I am not in the office today, but in email contact.
Thanks again for your help,
Kind Regards,

Read More

Abcam community

Verified customer

Asked on Jun 08 2012

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

Read More

Abcam Scientific Support

Answered on Jun 08 2012

Question

- 720 samples
- Interested in ab65354, ab83464, ab102530 and ab118970
- WIll use heparin containing plasma samples
- Activity kits

Read More

Abcam community

Verified customer

Asked on Jun 07 2012

Answer

Thank you for contacting us.

I can confirm that the kit ab65354 will detect all SODs (Cu/Zn, Mn and FeSOD).

If it is desired to measure SOD activity from cytosol and mitochondria separately,these can be separated by using ab65320 Mitochondria/Cytosol Fractionation Kit. SOD activity is then measured from the Mitochondria and Cytosol fractions separately.

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

Read More

Abcam Scientific Support

Answered on Jun 08 2012

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