Overview

  • Product name

    Fumarate Assay Kit
  • Detection method

    Colorimetric
  • Sample type

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

    Quantitative
  • Sensitivity

    > 1 nmol/well
  • Assay time

    0h 40m
  • Product overview

    Fumarate Assay Kit ab102516 provides a convenient tool for sensitive detection of Fumarate in biological samples.


    In the fumarate assay protocol, the fumarate enzyme mix recognizes fumarate as a specific substrate leading to proportional color development. The amount of fumarate can therefore be easily quantified using a colorimetric assay (λ = 450 nm). It can detect as low as 1 nmol of fumarate per well (20 µM).


    Fumurate assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 1 hr
    - analyze with a microplate reader

  • Notes

    Fumarate (HO2CCH=CHCO2H-) is an intermediate in the Kreb's cycle used by cells to metabolize food to form ATP. In the mammalian liver, Fumarate is also a product of the Urea cycle where its release in the cytosol leads to its conversion into malate and subsequently oxaloacetate while generating NADH in the cytosol.

  • Platform

    Microplate reader

Properties

Images

  • Fumurate Standard Curve
  • Fumarate measured in mouse tissue lysates (mg of extracted protein), background signal subtracted (duplicates +/- SD).

  • Fumarate measured in biological fluids, background signal subtracted (duplicates +/- SD).

  • Fumarate measured in cell culture medium and control medium, background signal subtracted (duplicates +/- SD).

  • Fumarate measured in cell lysates, background signal subtracted (duplicates +/- SD).

Protocols

References

This product has been referenced in:

  • Jin L  et al. The PLAG1-GDH1 Axis Promotes Anoikis Resistance and Tumor Metastasis through CamKK2-AMPK Signaling in LKB1-Deficient Lung Cancer. Mol Cell 69:87-99.e7 (2018). Read more (PubMed: 29249655) »
  • Kerins MJ  et al. Fumarate hydratase inactivation in hereditary leiomyomatosis and renal cell cancer is synthetic lethal with ferroptosis induction. Cancer Sci 109:2757-2766 (2018). Read more (PubMed: 29917289) »
See all 3 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Measurement of fumaric acid in stool

Good Good 4/5 (Ease of Use)
Abreviews
We investigated whether it was possible to measure fumaric acid in faeces. We tried several concentrations of faeces, and the results showed that it is possible to measure fumaric acid in faeces, although the measurement needs to be optimized.
Here is our protocol:
A Fumarate Detection Kit (Abcam 102516) was used for fumaric acid measurement in faeces. 40 mg of faeces was weighed and 100 ul Assay Buffer was added to the samples. Bead-beating (2 times 30 seconds) was performed, and afterwards centrifugation 10 minutes (13.000pm) for homogenization of the samples. The protein concentration of the supernatant was measured with the DC kit from Bradford Assay. Optimum protein concentration was determined testing various ug protein concentrations. After a couple try-outs, it appeared that 2.5 ul and 0.5 ul of the supernatant were the best concentrations to use in the assay. 2.5 ul and 0.5ul were both tested, and added to every well. Assay buffer was added to end up with a total concentration of 50 ul. Consequently, 100 ul reaction mix was added to each sample, which consisted of 90 ul Fumarate Assay Buffer, 8 ul Fumarate Developer and 2 ul Fumarate enzyme mix. After incubation of 60 minutes at 37°C, the absorbance was measured at 450 nm in a microplate reader.

Abcam user community

Verified customer

Submitted Jun 17 2016

Answer

As Fumarate is a chemical and not a protein, it's detection is not limited by species specificity as it would be the case if you would like to detect a protein.
In addition, as blood serum is blood plasma without clotting factors, and plasma has been tested as well for this kit, you should stick to protocol advise for serum.

Read More

Question
Answer

The final colour development should be pink but the reaction mix should not be pink to begin with. Only once you add this to your samples, will the reaction initiate and lead to the pink colour development.

Read More

Answer

Thank you for your enquiry.

I can confirm that any publications that we are aware of will be listed on the datasheet. We have a dedicated team who spend time reviewing the literature to find articles that have used our products.

However, in many cases, regrettably we do not have access to all the articles mentioning our products, or subscriptions to all the journals that publish them.

I'm sorry there are none to provide on this occasion for ab102516 (Fumarate Detection Kit).

But I am happy to confirm that this kit is tested and guaranteed for blood products such as serum and plasma.

Should you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you very much for your interest in our kit ab102516 for fumarate detection.

The laboratory confirmed that we haven’t tried this kit with microbial samples in-house and neither do we have any researcher feedback from such use. So although this kit can be theoretically used with microbial samples, practically, it might need some tweaking of the protocol at the user’s end to achieve the best results.

Therefore if you would like to test this kit on bacterial samples, I can offer a discount off a future purchase if you buy ab102516 now, test iton bacterial samplesand submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of this kit which we could takeoff your next order.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab102516 in bacterial samples. I will then send a discount code. This code must be issued before purchasing ab102516 so please wait for my reply before ordering.

2. Purchase ab102516 either by phone, fax, or online (www.abcam.com).

3. Test it in bacterial samples

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemedonyour next order and is worth the value of ab102516.The discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab102516 turns out to be unsuitable on bacterial samples, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up