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| Product Name | Lipid Peroxidation (MDA) Colorimetric Assay Kit |
| Catalog No. | CMTR-HMM-0060 |
| Description | A colorimetric assay for the quantification of malondialdehyde, a thiobarbituric acid reactive substance that serves as a biomarker of lipid peroxidation damage to cellular membranes. MDA reacts with thiobarbituric acid under acidic high-temperature conditions to form a pink MDA-TBA adduct measured spectrophotometrically at 532 nm. |
| Intended Use | Quantifying lipid peroxidation levels in cell and tissue samples for oxidative stress research, evaluation of antioxidant therapeutics, assessment of ischemia-reperfusion injury, and food quality analysis. |
| Principle / Technology | Unsaturated membrane fatty acids undergo peroxidation by reactive oxygen species to form lipid hydroperoxides, which decompose to produce MDA among other reactive aldehydes; MDA reacts with two molecules of thiobarbituric acid (TBA) at low pH and high temperature (95–100°C) to yield the pink chromogen MDA-(TBA)2, with maximal absorbance at 532 nm. |
| Detection Method | Colorimetric absorbance at 532 nm using spectrophotometer or microplate reader; also detectable by fluorescence (Ex/Em 532/553 nm) for increased sensitivity |
| Sample Type | Cell and tissue homogenates, serum and plasma, urine, tissue culture supernatant, subcellular fractions (microsomes, mitochondria), food and biological oils |
| Performance Range / Specifications | MDA standard range: 0.5–50 μM (linear 0–100 nmol range); incubation of TBA reaction at 95°C for 60 minutes; color stable for approximately 2 hours after reaction completion; absorbance 0.05–2.0 OD at 532 nm |
| Sensitivity / LOD | Detection limit: approximately 0.1 μM MDA in sample with 100 μL sample volume; equivalents to approximately 1 μg/mL in tissue homogenate at 1:10 dilution |
| Specificity | TBA reaction detects MDA with high specificity, but other aldehydes present under strong oxidative stress conditions (4-hydroxynonenal, acrolein) can contribute minor absorbance; specificity improved by measuring the MDA-specific absorbance at 532 nm and subtracting non-specific background at 450 nm and 600 nm per the 3-wavelength correction method |
| Reaction Conditions / Protocol | Prepare cell or tissue lysates (homogenize in PBS or RIPA buffer with antioxidant BHT); add 100 μL sample or MDA standard to tubes; add 200 μL TBA Color Development Solution; incubate at 95–100°C for 60 minutes in a heating block; cool to room temperature in ice bath (10 minutes); centrifuge at 3,000 × g for 5 minutes to remove turbidity; transfer supernatant and measure absorbance at 532 nm; for low-level MDA samples, extract the MDA-TBA chromogen into n-butanol and measure the organic phase absorbance |
| Components / Formulation | MDA Standard (1,1,3,3-tetramethoxypropane, a stable MDA precursor), TBA Reagent (thiobarbituric acid, 0.6% w/v), SDS Solution, Acetic Acid Solution (pH 3.5), BHT (butylated hydroxytoluene, antioxidant to prevent further lipid oxidation), n-butanol for extraction protocol |
| Storage Conditions | MDA standard: -20°C in desiccated condition; TBA reagent: room temperature protected from light, prepare fresh working solution; acetic acid and SDS solutions: room temperature, 24 months |
| Shelf Life | 12 months from date of manufacture for chemical standards; 24 months for buffer components |
| Package Specifications | 100 assays, 500 assays (microcentrifuge tube format or 96-well plate format) |
| Product Form | Liquid and lyophilized chemical reagents for assembling the TBA reaction |
| Quality Control | Each lot verified with MDA standard curve (R2 ≥0.99); positive control using H2O2-treated erythrocyte lysate; 3-wavelength correction method validated to remove non-specific absorbance contributions |
| Key Features | TBA method is the most extensively published lipid peroxidation assay providing historical comparability; stable MDA standard (tetramethoxypropane) yields consistent calibration; colorimetric detection accessible to any spectrophotometer-equipped laboratory; 3-wavelength correction improves analytical specificity |
For research use only, not for clinical use.
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