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.
Submaximal exercise training, more than dietary selenium supplementation, improves antioxidant status and ameliorates exercise-induced oxidative damage to skeletal muscle in young equine athletes
Yong Zhi Foo, Leigh W. Simmons, Gillian Rhodes
Journal of Animal Science ,  Vol. 95 No. 2, p. 657-670

View Abstract

Time-Dependent Toxicity Responses in Daphnia magna Exposed to CuO and ZnO Nanoparticles
Soyoun KimPalas SamantaJisu YooWoo-Keun KimJinho Jung
Bulletin of Environmental Contamination and Toxicology ,  April 2017, Volume 98, Issue 4, pp 502–507

View Abstract

Chronic Unpredictable Stress Induces Renal Damage In Rats By Oxidative Stress Provoked Apoptosis and Altering the Function of Na+/K+-ATPase
I Bin-Jaliah
Pak J Physiol ,  2016;12(3)

View Abstract

Dose-dependent effects of cisplatin on the severity of testicular injury in Sprague Dawley rats: reactive oxygen species and endoplasmic reticulum stress
Kiran Kumar Soni, Hye Kyung Kim, Bo Ram Choi, Keshab Kumar Karna, Jae Hyung You, Jai Seong Cha, Yu Seob Shin, Sung Won Lee, Chul Young Kim, and Jong Kwan Park
Drug Design, Development and Therapy ,  2016; 10: 3959–3968

View Abstract

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

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

Gastroprotective Effect of Ethanolic Extract of Curcuma xanthorrhiza Leaf against Ethanol-Induced Gastric Mucosal Lesions in Sprague-Dawley Rats
Nurhidayah Ab. Rahim, Pouya Hassandarvish, Shahram Golbabapour, Salmah Ismail, Saad Tayyab, and Mahmood Ameen Abdulla
BioMed Research International Volume 2014; Article ID 416409, 10 pages

Link to Abstract

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