Malondialdehyde (MDA) Assay Kit: 200 Tests

NWLSS | Northwest Life Science Specialties, LLC.
Malondialdehyde (MDA) Assay
NWK-MDA01 $315.00 each

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A colormetric assay kit for detection of Malondialdehyde (MDA) or Thiobarbituric Acid Reactive Substances (TBARS) in multiple species and sample types. Designed for simplicity and affordability, this assay utilizes various improvements to provide the most dependable data among commercial assays of this type.

Advantages of the NWLSS™ Malondialdehyde Assay:
  • BHT and EDTA are added to the sample and reaction mixture to minimize artifact oxidation.
  • Reduced reaction temperature minimizes the decomposition of lipid hydroperoxides.
  • Optimized reaction pH facilitates hydrolysis of MDA-protein adducts for better recovery of malondialdehyde (MDA).
  • Cleaner output through optimized data reduction using dual wavelength or 3rd derivative analysis of spectroscopic scan data.

Introduction

Lipid peroxidation has been established as a major mechanism of cellular injury in many biological systems of plant and animal origin. The mechanism involves a process whereby unsaturated lipids are oxidized to form additional radical species as well as toxic by-products that can be harmful to the host system. Polyunsaturated lipids are especially susceptible to this type of damage when in an oxidizing environment and they can react to form lipid peroxides.

Lipid peroxides are themselves unstable, and undergo aditional decomposition to form a complex series of compounds including reactive carbonyl compounds. Polyunsaturated fatty acid peroxides further react to form malonaldehyde (MDA).

MDA can be found in most biological samples including foodstuffs, serum, plasma, tissues and urine, as a result of lipid peroxidation, and has become one of the most widely reported analytes for the purpose of estimating oxidative stress effects on lipids.

Test Principle

This assay is based on the reaction of malondialdehyde (MDA) with thiobarbituric acid (TBA); forming a MDA-TBA2 adduct that absorbs strongly at 532 nm.

Thiobarbituric acid reactive substances (TBARS) Reaction
MDA - TBA Reaction

This reaction is the most popular method for estimating MDA in biological samples. However, interference can be a significant problem in some biological samples if not dealt with appropriately. Our assay provides the most accurate and efficient means of dealing with elevated backgrounds commonly associated with the TBARS reaction.

Can the NWK-MDA01 assay be formatted for use in the 96-well microplate format?

Yes the reaction can be scaled down and the reactin product read in a 96 well plate. However, you may lose some sensitivity due to the reduced path length associated with the lower volume necessary for reading on a plate. Keep in mind that "normal" human plasma or serum is expected to have less than 1 uM...usually around 0.25 uM MDA. You may scale as necessary by simply keeping the ratio of sample and reagents to one another the same. The reaction should still be run in a microcentrifuge tube allowing for centrifugation and subsequent transfer to a microplate for reading.

Is there any way to get a good measurement for MDA when testing cloudy samples?

Regarding the issue of turbidity, occasionally, it becomes necessary to clean up samples that appear cloudy. For this, we do have a back extraction protocol online (see http://www.nwlifescience.com/bext/ ) that may be useful to you. The protocol was originally created to remove excess hemoglobin from reacted samples but can also be useful to remove other potentially interfering substances.

I am seeing precipitate after adding the 60 minute incubation at step 6. Should I be concerned?

Regarding the precipitate seen at step # 6: I believe that this is an acid induced precipitation of proteins which is why there is a required centrifugation at step # 7 and subsequent transfer of the reaction mixture to either cuvette or plate for reading at step # 8.

Can MDA be detected in urine using the NWK-MDA01 kit?

Yes. However, the user should understand that the recovery of MDA (TMOP) in urine is roughly 50-70%. NWLSS data shows that urea can interfere with the assay and may be a potential cause for the lower recovery. That being said, many customers have used the kit to detect MDA in urine over the years. Here are a few citations wherein we know that product NWK-MDA01 has been used to test MDA in urine:

I want to compare lipid peroxidation in two different cell culture populations. Can I measure MDA in the cell medium or do I need to assay a cell homogenate?

We recommend homogenizing the cells in the Assay Buffer provided, centrifuging to clarify then testing the clarified supernatant.

Can I use Chloroform (CHCl3) to extract my samples similar to the butanol based method shown at www.nwlifescience.com/bext/

We have compared CHCl3 and BuOH extractions. We found that with CHCl3 there is incomplete partitioning of heme into the organic phase which results in some heme in the aqueous phase containing the MDA-TBA2 assay product. Excess heme in the MDA-TBA2 containing aqueous phase could result in false positives. Alkaline BuOH extraction does achieve 100% separation of heme from MDA-TBA2.
Sudan III Azo Dye: Oxidative Stress with Possible Geno and Hepatotoxic Effects in Male Rats
Dildar Konukoglu, Sinem Fırtına, Gokhan Erkol and I. Murat Bolayırlı
International Journal of Science and Research (IJSR) ,  Volume 5 Issue 10, October 2016

View Abstract

Removal of harmful alga, Chattonella marina, by recyclable natural magnetic sphalerite
Bo Wanga, Dan Wua, Ka Him Chua, Liqun Yeb, Ho Yin Yipa, Zhonghua Caic, Po Keung Wonga
Journal of Hazardous Materials,  Available online 9 November 2016

View Abstract

β-Caryophyllene, a phytocannabinoid attenuates oxidative stress, neuroinflammation, glial activation, and salvages dopaminergic neurons in a rat model of Parkinson disease
Eunhye Bae, Palas Samanta, Jisu Yoo, Jinho Jung
Molecular and Cellular Biochemistry ,  July 2016, Volume 418, Issue 1, pp 59–70

View Abstract

Effects of multigenerational exposure to elevated temperature on reproduction, oxidative stress, and Cu toxicity in Daphnia magna
Eunhye Bae, Palas Samanta, Jisu Yoo, Jinho Jung
Ecotoxicology and Environmental Safety ,  Volume 132, October 2016, Pages 366–371

View Abstract

Impact of antioxidants on seminal vesicles function and fertilizing potential in diabetic rats
Panagiota Tsounapi, Masashi Honda, Fotios Dimitriadis, Bunya Kawamoto, Katsuya Hikita, Kuniyasu Muraoka, Motoaki Saito, Nikolaos Sofikitis, Atsushi Takenaka
Asian Journal of Andrology ,  Availale Online October 14, 2016.

View Abstract

Browse all Citations

For Research Use Only

Catalog Number: NWK-MDA01
Format: TBA Based Colorimetric
Sample Requirements: Tissue homogenates, cell lysates and plasma
Specificity: Primary specificity is for malondialdehyde (MDA) when using advanced data reduction and/or back extraction techniques. Basic Thiobarbituric acid reactive substances (TBARS) are detected when making single wavelength 532nm measurements.
Sensitivity: 0.1 µM MDA in the sample
Standard Range: 1.0 - 4.0 mM
Storage and Stability: 12 months from date of manufacture when stored at 2-8 C
Kit Contents: 5 X TBA in powder form
5 X H3PO4; ready to use
5 X TMOP Calibrators; ready to use
1 x Butylated Hydroxytoluene (BHT; ready to use)
1 x Assay Buffer
Random Citation | NWK-MDA01

Long-Term Moderate Dose Exogenous Erythropoietin Treatment Protects from Intermittent Hypoxia-Induced Spatial Learning Deficits and Hippocampal Oxidative Stress in Young Rats
Jobran M. Al-Qahtani, Basel A. Abdel-Wahab, Samy M. Abd El-Aziz
Neurochemical Research January, 2014; Volume 39, Issue 1, pp 161-171

Link to Abstract

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