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

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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.

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Verified customer

Submitted Jun 17 2016

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