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

Reviews (1)Q&A (14)References (545)
MDA assay standard curve
  • MDA assay standard curve
  • Lipid Peroxidation Assay performed on mouse sciatic nerve samples
  • Lipid Peroxidation measured with MDA assay in Fabry patients and healthy controls
  • Functional Studies - Lipid Peroxidation (MDA) Assay Kit (ab118970)

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric/Fluorometric
  • Platform: Microplate reader
  • Assay time: 1 hr 20 min
  • Sample type: Cell culture extracts, Plasma, Tissue Extracts, Urine (UTI)
  • Sensitivity: 0.1 nmol/well

製品の概要

  • 製品名

    Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric)
    Lipid Peroxidation キット 製品一覧

  • 検出方法

    Colorimetric/Fluorometric

  • サンプルの種類

    Plasma, Cell culture extracts, Tissue Extracts, Urine (UTI)

  • アッセイタイプ

    Quantitative

  • 検出感度

    > 0.1 nmol/well

  • 全工程の試験時間

    1h 20m

  • 製品の概要

    Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) (ab118970) provides a convenient tool for sensitive detection of malondialdehyde (MDA).


    How the assay works
    In the lipid peroxidation assay protocol, 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.


    Lipid peroxidation assay protocol summary


    - Add TBA solution to samples and standards, incubate at 95°C for 60 min, cool in ice bath for 10 min
    - Transfer to wells of microplate
    - Analyze with microplate reader


    For higher sensitivity, precipitate with n-butanol, centrifuge, dry and resuspend pellet before analysis.


    Related MDA assay products
    For an alternative MDA assay, without the heating steps required in the TBARS assay, try MDA assay ab233471.


    Getting the best performance from ab118970
    Technical Note: MDA is unlikely to be detectable in healthy urine but may be elevated in the presence of infection or disease of the urinary tract.


    How other researchers are using Lipid Peroxidation Assay Kit ab118970
    The MDA/TBARs assay kit has been used in publications in a variety of sample types, including:


    - Human: serum1, hippocampal primary cell extracts2, A375 cultured cell lysates3, plasma and platelet samples4
    - Mouse: neuronal cell lysates5, heart tissue extract6, plasma7, cell extracts8
    - Rat: hippocampal tissue extracts9, cardiomyocyte extracts of cultured cells10, lung lysates11
    - Pig: serum12


    References: 1 - Shen J et al. 2018, 2 - Wang Q et al. 2019, 3 - Luo M et al. 2018, 4 - Mustafa AG et al. 2018, 5 - Murphy K et al. 2018, 6 - Guan F et al. 2019, 7 - Costa CRC et al. 2018, 8 - Eleftheriadis T et al. 2019, 9 - Malekiyan et al. 2019, 10 - Zhou Z et al. 2018, 11 - Li L et al. 2018, 12 - Lee SE and Kang KS 2019


    Related and recommended products
    Also see the popular 4-HNE Assay Kit ab238538 as an alternative marker of lipid peroxidation and oxidative stress.


    Iron Assay Kit ab83366 is often used in combination with Lipid Peroxidation (MDA) Assay Kit ab118970 and GSH/ GSSG Assay Kit ab239709 in the study of ferroptosis.


    Background information
    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. Lipid peroxidation forms reactive aldehydes such as malondialdehyde (MDA) and 4-hydroxynonenal (4- HNE) as natural bi-products. Malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) are often used as markers of lipid peroxidation, and to assay for oxidative damage / oxidative stress.


    The MDA assay is also refered to as a TBARS assay. The TBARS assay (thiobarbituric acid reactive substance assay) is used to measure lipid peroxidation in cell and tissue extracts, and biological fluids. The TBARS assay detects the level of MDA (malondialdehyde), the major lipid oxidation product, and also some minor related compounds. It is often considered a good index of the level of oxidative stress in a biological sample.


    Other notes
    This product is manufactured by BioVision, an Abcam company and was previously called K739 Lipid Peroxidation (MDA) Colorimetric/Fluorometric Assay Kit. K739-100 is the same size as the 100 test size of ab118970.

  • 特記事項

    The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.

  • 試験プラットフォーム

    Microplate reader

製品の特性

画像

  • MDA assay standard curve
    MDA assay standard curve

    Typical MDA standard calibration curve using fluorometric reading.

  • MDA assay standard curve
    MDA assay standard curve

    Typical MDA standard calibration curve using colorimetric reading.

  • Lipid Peroxidation Assay performed on mouse sciatic nerve samples
    Lipid Peroxidation Assay performed on mouse sciatic nerve samplesImage courtesy of Hichor et al. Sci Rep. 2018; 8: 2524. doi: 10.1038/s41598-018-20980-3. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.
    Hichor M et al. used the TBARS assay /  MDA assay ab118970 to study the role of LXRs in the regulation of oxidative stress in peripheral  nerves.
     
    They identified that in sciatic nerves in LXR knockout mice (LXRdKO), the MDA concentration was significantly increased, and that this was corrected by the treatment of mice with the anti-oxidant ROS scavenger N-acetylcysteine (NAC).
  • Lipid Peroxidation measured with MDA assay in Fabry patients and healthy controls
    Lipid Peroxidation measured with MDA assay in Fabry patients and healthy controlsImage courtesy of Ravarotto V et al. PLoS One. 2018; 13(9): e0204618. doi: 10.1371/journal.pone.0204618. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

    Ravarotto V et al. used Lipid Peroxidation Assay Kit ab118970 to assess oxidative stress in Fabry disease. They identified that MDA levels are higher in Fabry patients, indicating higher levels of oxidative stress.

    The MDA concentration was measured in plasma from Fabry patients compared to healthy control patients. Data are shown ±SEM. *: p = 0.01.

  • Functional Studies - Lipid Peroxidation (MDA) Assay Kit (ab118970)
    Functional Studies - Lipid Peroxidation (MDA) Assay Kit (ab118970)

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

参考文献 (545)

ab118970 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab118970 は 545 報の論文で使用されています。

  • Fouad H  et al. Use of Mesenchymal Stem Cells in Experimental Ovarian Damage. Curr Stem Cell Res Ther 19:725-734 (2024). PubMed: 37448361
  • Bi R  et al. Butyrate enhances erastin-induced ferroptosis of lung cancer cells via modulating the ATF3/SLC7A11 pathway. Environ Toxicol 39:529-538 (2024). PubMed: 37341073
  • Ji M  et al. Vinpocetine improves dyskinesia in Parkinson's disease rats by reducing oxidative stress and activating the Wnt/β-catenin signaling pathway. Chem Biol Drug Des 103:e14358 (2024). PubMed: 37749299
  • Qin H  et al. Tanshinone IIA ameliorates cisplatin-induced toxicology and cisplatin resistance via regulating SLC7A11 expression. Environ Toxicol 39:1429-1441 (2024). PubMed: 37987512
  • Niu T  et al. Porous Se@SiO2 nanospheres alleviate diabetic retinopathy by inhibiting excess lipid peroxidation and inflammation. Mol Med 30:24 (2024). PubMed: 38321393
View all Publications for this product

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1-10 of 15 Abreviews or Q&A

Good kit for alternative samples

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We used this as an alternative to a TBARs assay on our in vitro digestion supernatant. We got good readings, however, there was some unexpected color formation (brown/yellow) in the test wells compared to the standard wells (pink). We are not sure if this altered the results at all, but our values are in the expected range. We did have to dilute our sample.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jan 17 2023

Question

I have a few questions about this kit:
1. Can I mix more than 10ul plasma with 500ul sulfuric acid? I am trying to boost signal by adding more concentrated sample.
2. It seems the standard curve produced by the kit is a good one but I see no signal from plasma samples. I have followed the protocol exactly. Do you have any ideas why one would not see any signal with plasma?
3. After heating and cooling the samples, do I need to run the assay immediately or can the cooled samples be stored? If storage is possible, what do you recommend?

Read More
Answer

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 fluorometric method might provide 10 fold increased sensitivity to detect this. It is possible in that case to not register any OD or see background OD with samples.
A3: For best results we do not recommend storing samples after 95C heating and cooling. Lipid binding to the walls of the tube can happen upon storage and this can skew the data.

Read More

Question

The TBA is not going into solution in the glacial acetic acid.

Read More
Answer

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 precipitate, use 600 ul as directed and any particle in suspension will dissolve at 95C when incubated for 60 mins.

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Question

How are the nmol standard curve units converted to units of uM concentrations and micrograms?

Read More
Answer

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 values you obtained.

The standard you receive in the kit is 4.17M. Dilute 10ul to 0.1M, then 2mM. Standards for the colorimetric assay are 0, 2, 4, 6, 8, 10 ul of the 2mM solution, per tube. The amounts per tube are specified in the protocol and the data as nanomoles (nmol), not nanomolar concentrations (nM).

For example, the 2mM standard solution contains: 2 millimoles per liter, = 2 nanomoles per microliter. The standard that contains 2 ul contains 4 nanomoles.

The molar concentration of this standard, after bringing the volume to 200ul (which is also the volume of the experimental samples), is 4 nanomoles per 200ul, = 20,000 nanomoles per liter (4 x 5000 nanomoles per 200 x 5000 ul) = 20 micromoles (umol) per liter, which is a 20 micromolar (uM) solution.

So, you are comparing your samples to MDA standards that are concentrations of 20, 40, 60, 80, and 100 uM .

The molecular weight of MDA is 72g/L. (One mole weighs 72g).

A 20 uM solution contains 1440 ug per liter. (72/1,000,000 x 20).

In 200 ul, you will have: 1440/5,000 ug, = 0.288 ug.

So, the standard tubes contain: 0.288, 0.432, 0.576, 0.72, and 1.44 ug per tube.

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Question

can you tell me where in the protocol I can stop the assay and how the samples should be stored at that point?

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Answer

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 the protocol.

Read More

Question

Appropriate sample types include Cell culture supernatant, Plasma, Other biological fluids andTissue Extracts. I would like to know I can use this kit for checking MDA in serum samples of human.

Read More
Answer

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, to try the procedure provided for plasma to start with and optimize from there.

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Question

We don't have 532 nm of colorimetric spectro filter in my lab, we only have the 520nm. And so, is it ok if I use 520nm? would it effect the accuracy of the measurement significantly?

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Answer

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.

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Question

Would heparin treated plasma samples be compatible with this kit?

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Answer

Yes, heparin should be fine to use.

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Question

The standards work well but I see precipitation in my sample after heating at 95 degrees C for 60 min. Samples are cell extracts.
1. Is this normal?
2. How should this be addressed?

Read More
Answer

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

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Question

At the end of the protocol there is a note about the use of n-butanol for more sensitivity. I must add the n-butanol to the 800 µl reaction mixture. Do I have to add the n-butanol also to the standard or only the samples?

Read More
Answer

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.

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1-10 of 15 Abreviews or Q&A

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