Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) (ab118970)

Overview

  • Product name
    Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric)
  • Detection method
    Colorimetric/Fluorometric
  • Sample type
    Plasma, Cell culture extracts, Tissue Extracts
  • Assay type
    Quantitative
  • Sensitivity
    > 0.1 nmol/well
  • Product overview

    Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) (ab118970) provides a convenient tool for sensitive detection of the malondialdehyde (MDA) present in a variety of samples. MDA, together with 4-hydroxynonenal (4-HNE), is a natural bi-product of lipid peroxidation and its quantification is generally used as marker for lipid peroxidation. The MDA in the sample reacts with thiobarbituric acid (TBA) to generate a MDA-TBA adduct. The MDA-TBA adduct can be easily quantified colorimetrically (OD = 532 nm) or fluorometrically (Ex/Em = 532/553 nm). This assay detects MDA levels as low as 1 nmol/well colorimetrically and 0.1 nmol/well fluorometrically.


    The MDA assay is also refered to as a TBARS assay.


    Visit our FAQs page for tips and troubleshooting.


    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • Notes

    Lipid peroxidation refers to the oxidative degradation of lipids. In this process free radicals take electrons from the lipids (generally in cell membranes), resulting in cell damage. Quantification of lipid peroxidation is essential to assess oxidative stress in pathophysiological processes. Lipid peroxidation forms reactive aldehydes such as malondialdehyde (MDA) and 4-hydroxynonenal (4- HNE) as natural bi-products. Measuring the end products of lipid peroxidation is one of the most widely accepted assays for oxidative damage.

  • Platform
    Microplate

Properties

Images

  • 10 mg of tissue were homogenized on ice in 300 μL of MDA lysis buffer, then centrifuged (13,000 × g, 10 min) to remove insoluble materials. 10 μL of plasma were mixed with 500 μL of 42 mM H2SO4 and 125 μL of phosphotungstic acid solution at room temperature for 5 min. After centrifuging (13,000 × g, 3 min), the pellet was re-suspended on ice with 100 μL of double-distilled H2O. Then, 200 μL of solution and 600 μL of 2-thiobarbituric acid solution were incubated at 95°C for 60 min, before cooling to room temperature in the ice bath for 10 min. The intensity of absorbance at 532 nm was proportional to the MDA level.

  • Typical MDA standard calibration curve using colorimetric reading.

  • Typical MDA standard calibration curve using fluorometric reading.

  • Measurement of MDA in human plasma (20 µl) and rat liver lysate (10 mg).

Protocols

References

This product has been referenced in:
  • Hichor M  et al. Liver X Receptor exerts a protective effect against the oxidative stress in the peripheral nerve. Sci Rep 8:2524 (2018). Read more (PubMed: 29410501) »
  • Hai B  et al. Delivery of Sonic Hedgehog Gene Repressed Irradiation-induced Cellular Senescence in Salivary Glands by Promoting DNA Repair and Reducing Oxidative Stress. Theranostics 8:1159-1167 (2018). Read more (PubMed: 29464006) »

See all 44 Publications for this product

Customer reviews and Q&As

The MDA kit should be compatible with RIPA but the concentration of SDS needs to be less than 5mM.

Below please find answers to your questions:

A1: We have found that the 10 ul plasma worked well. How much more plasma is used depends on the protein content.
A2: If there is low amounts of MDA adduct formed in plasma samples, then the flu...

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Sonication in a RT water bath can be used if needed. You can also dissolve TBA in 15 ml of 50% glacial acetic acid in water. Then fill the volume with water to make 25ml. Mix well. This should help in dissolving the TBA. If there is still little precip...

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Here is my conversion from nanomoles to micromolar concentrations and micrograms of MDA for the units that are used for the X-axis of the data on the datasheet. Please let me know if this does not answer you question or if you have concerns about the v...

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We do not recommend that the protocol be stopped for later continuation at any step for this assay. If you cannot perform the whole protocol on the same day, I would recommend storing the samples initially at -80°C for later processing according to...

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I am sorry to confirm this kit has not been tested on serum samples yet. Since it has not been used for analysis of serum, I am sorry we unfortunately do not have any procedure or tips for such use. I can recommend that as plasma samples are similar, t...

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I can confirm you would be able to use the 520 nm wavelength for detection, provided the instrument's bandwidth of detection range is +/- 12. If not, measuring at 520 nm will reduce the efficiency of detection at least slightly.

Yes, heparin should be fine to use.

Centrifuge the samples and take only the clear supernatant for the next step.

I can confirm that you will need to use the n-butanol for the standards as well, since it helps in isolating the MDA-TBA adduct.

1-10 of 15 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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