Lipid peroxidation mda assay kit

Manufactured by Merck Group

Sourced in United States, Germany, Sao Tome and Principe

The Lipid Peroxidation (MDA) Assay Kit is a laboratory tool used to measure the level of malondialdehyde (MDA), a byproduct of lipid peroxidation. It provides a quantitative assessment of oxidative stress in biological samples.

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Close spelling variants of this product

Lipid Peroxidation MDA Assay Lipid peroxidation kit MDA kit

67 protocols using lipid peroxidation mda assay kit

Quantifying Cellular Lipid Peroxidation

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Malondialdehyde (MDA) concentration in cellular extracts was assessed with the Lipid peroxidation (MDA) assay kit from Merck (Darmstadt, Germany, MAK085) according to manufacturer’s instructions.

Vermonden P., Vancoppenolle M., Dierge E., Mignolet E., Cuvelier G., Knoops B., Page M., Debier C., Feron O, & Larondelle Y. (2021). Punicic Acid Triggers Ferroptotic Cell Death in Carcinoma Cells. Nutrients, 13(8), 2751.

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Lipid Peroxidation in Muscle Tissue

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Lipid peroxidation in muscle tissue was assessed using a Lipid Peroxidation (MDA) Assay Kit (Merck, Darmstadt, Germany), according to the manufacturer’s instructions. Briefly, 10 mg of tissue was homogenized on ice in MDA Lysis Buffer containing BHT and centrifuged. The supernatant was mixed with TBA solution and incubated at 95 °C for 60 min. Absorbance was measured at 532 nm, and the concentration of malondialdehyde in samples was extrapolated from its standard curve.

Mićić B., Djordjevic A., Veličković N., Kovačević S., Martić T., Macut D, & Vojnović Milutinović D. (2023). AMPK Activation as a Protective Mechanism to Restrain Oxidative Stress in the Insulin-Resistant State in Skeletal Muscle of Rat Model of PCOS Subjected to Postnatal Overfeeding. Biomedicines, 11(6), 1586.

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Oxidative Stress Biomarkers Determination

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Superoxide Dismutase Determination Kit (SOD, 19160-1KT-F), Lipid Peroxidation (MDA) Assay Kit (MAK085-1KT), Glutathione Peroxidase Cellular Activity Assay Kit (GPx, CGP1-1KT), and Total Protein Kit, Micro Lowry, Peterson’s Modification (TP0300-1KT) were purchased from Merck, Germany. Physiological saline (0.90% NaCl) was purchased from Hemofarm (Romania). Ascorbic acid (Vit. C) was purchased from a local pharmacy in order to do a study in more related situations and conditions. The Vit. C was in a liquid form, soluble in water, and designated for children as a supplement. The composition of this supplement was acid ascorbic, glycerin, propylene glycol, and orange flavor. The supplement was diluted the desired concentration was obtained. For fipronil and pyriproxyfen (1:1), we used a common insecticide that is soluble in water and contains these two compounds with a quality certificate and it was purchased from a local store. Every 1.5 mL of this insecticide had the following composition: 67.5 mg of fipronil, 67.5 mg of pyriproxyfen, 0.3 mg of butylated hydroxyanisole, 0.15 mg of butylhydroxytoluene, and 60 mg of benzyl alcohol.

Robea M.A., Jijie R., Nicoara M., Plavan G., Ciobica A.S., Solcan C., Audira G., Hsiao C.D, & Strungaru S.A. (2020). Vitamin C Attenuates Oxidative Stress and Behavioral Abnormalities Triggered by Fipronil and Pyriproxyfen Insecticide Chronic Exposure on Zebrafish Juvenile. Antioxidants, 9(10), 944.

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Measuring Cellular and Serum MDA Levels

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Malondialdehyde concentration in cellular extracts was determined with the Lipid Peroxidation (MDA) Assay Kit from Merck (MAK085), according to manufacturer's instructions. Malondialdehyde concentration in mouse serum was determined with the Mouse malondialdehyde (MDA) ELISA kit from Mybiosource (MBS269473), according to manufacturer's instructions.

Dierge E., Debock E., Guilbaud C., Corbet C., Mignolet E., Mignard L., Bastien E., Dessy C., Larondelle Y, & Feron O. (2021). Peroxidation of n-3 and n-6 polyunsaturated fatty acids in the acidic tumor environment leads to ferroptosis-mediated anticancer effects. Cell metabolism, 33(8).

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MDA Levels Quantification in Brain Tissue

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The MDA levels in the brain sample were assayed using a Lipid Peroxidation (MDA) Assay Kit (Sigma-Aldrich Co.) according to the manufacturer's protocols. Briefly, 45 mg cortex and 5 mg hippocampus tissue from each group of mice were homogenized in MDA lysis buffer containing butylhydroxytoluene (BHT), after which the homogenates were stored at -20℃ until analysis. The sample or standards and TBA solution (70 mM thiobarbituric acid and 5M glacial acetic acid) were incubated at 95℃ for 60 min, then cooled to room temperature in an ice bath for 10 min, after which the reaction absorbance at 532 nm was read using a Versa max plate reader (Molecular Devices).

Koh E.K., Yun W.B., Kim J.E., Song S.H., Sung J.E., Lee H.A., Seo E.J., Jee S.W., Bae C.J, & Hwang D.Y. (2016). Beneficial effect of diosgenin as a stimulator of NGF on the brain with neuronal damage induced by Aβ-42 accumulation and neurotoxicant injection. Laboratory Animal Research, 32(2), 105-115.

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Evaluating Oxidative Stress in HMEC-1

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To determine whether PM2.5 affected the ROS levels of HMEC-1, a 2′,7′-dichlorofluorescin diacetate (DCFDA) kit (Sigma-Aldrich; Merck KGaA) was used. Curcumin/PM2.5 treated HMEC-1 (2×10⁵ cells/well) were added to DCFDA (20 µM) for 20 min, washed with PBS and then the ROS expression was analyzed by flow cytometry All experiments were repeated three times.
The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity of HMEC-1 were detected by a Lipid Peroxidation (MDA) assay kit (Sigma-Aldrich; Merck KGaA) and SOD assay kit (Sigma-Aldrich; Merck KGaA), respectively.

Shi J., Deng H, & Zhang M. (2017). Curcumin pretreatment protects against PM2.5-induced oxidized low-density lipoprotein-mediated oxidative stress and inflammation in human microvascular endothelial cells. Molecular Medicine Reports, 16(3), 2588-2594.

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Intracellular Iron and Lipid ROS Detection

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The relative level of intracellular iron was detected by an Iron Assay Kit (Abcam, USA) following manufacturer’s protocol. The lipid ROS was detected by using C11-BODIPY (Invitrogen) staining. In brief, cells were planted on 12-well plates and incubated overnight. After indicated incubation, cells were collected and resuspended in 200 μL PBS, stained with C11-BODIPY (5 μM) at 37°C for 30 minutes. Samples were examined by flow cytometer (BD, USA). The level of malondialdehyde (MDA) was assessed by using a lipid peroxidation (MDA) assay kit (Sigma). Cells were lysed by MDA lysis buffer on ice, followed by centrifugation at 13,000 × g for 5 minutes. The absorbance values were detected at 532 nm by a spectrophotometer (BD Biosciences).

He G.N., Bao N.R., Wang S., Xi M., Zhang T.H, & Chen F.S. (2021). Ketamine Induces Ferroptosis of Liver Cancer Cells by Targeting lncRNA PVT1/miR-214-3p/GPX4. Drug Design, Development and Therapy, 15, 3965-3978.

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Measuring Lung MDA Levels

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The malondialdehyde (MDA) level in lung tissues and cells was assessed using the Lipid Peroxidation (MDA) Assay Kit (Sigma, Cat #: MAK085), according to the manufacturer's instructions.

Lin Y., Cai J., Huang D., Zhou B., Luo Z., Yu S, & Lu J. (2021). Effects of dexmedetomidine on the expression profile of tsRNAs in LPS‐induced acute lung injury. Journal of Clinical Laboratory Analysis, 36(1), e24115.

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Quantifying Oxidative Stress in HUVECs

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A total of 3 × 10⁴ treated HUVECs were collected to determine the level of oxidative injury. The reactive oxygen species (ROS) and malondialdehyde (MDA) levels were detected according to the instruction books of the Total Reactive Oxygen Species Assay Kit (Invitrogen, Thermo Fisher Scientific, Waltham, MA) and the Lipid Peroxidation (MDA) Assay Kit (Sigma).

Mei R., Wu M, & Ren F. (2022). Knockdown of circ_0002194 protects against oxidized low-density lipoprotein-induced cell damage via the regulation of the miR-637/PACS2 axis in human vascular endothelial cells. Interactive Cardiovascular and Thoracic Surgery, 35(4), ivac210.

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Saliva MDA Quantification Protocol

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We measured saliva MDA content using a commercially available MDA Assay Kit (Sigma-Aldrich, Lipid Peroxidation MDA Assay Kit) following the instructions provided by the supplier; MDA levels are expressed as OS levels [24 (link)]. In brief, a mixture of 100 μL saliva fluid is diluted with 200 μL buffer provided in the kit. The saliva sample in buffer and Thiobarbituric acid (TBA) solution, 600 μL each, were mixed thoroughly and incubated at 95 °C for 60 min. Of the reaction mixture, after cooling in ice, 150 μL was transferred to a 96 well microplate in duplicates and absorbance was measured flurometrically (Epoch Microplate, BioTek Instrument, Vermont, USA) by measuring fluorescence intensity (λ_ex = 532/ λ_em = 553). The MDA levels in the saliva were calculated by the MDA standard provided in the kit with the help of standard curve and regression equation (R² = 0.9903 and y = 728.95x + 111.6).

Wagle M., Basnet P., Vartun Å., Trovik T.A, & Acharya G. (2020). Oxidative stress levels and oral bacterial milieu in the saliva from pregnant vs. non-pregnant women. BMC Oral Health, 20, 245.

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