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Malondialdehyde (MDA) Colorimetric Assay Kit (TBA Method)
SKU: E-BC-K025-M-500
Malondialdehyde (MDA) Colorimetric Assay Kit (TBA Method)
| SKU # | E-BC-K025-M |
| Detection Instrument | Microplate reader (530-540 nm, optimum wavelength: 532 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | MDA |
| Sample Type | Serum, plasma, animal tissue |
| Sensitivity | 1.13 μmol/L |
| Detection Range | 2.92-40 μmol/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 65 min |
| Precision | Average inter-assay CV: 7.200% | Average intra-assay CV: 4.100% |
| Other instruments required | Micropipettor, Vortex mixer, Incubator, Centrifuge, Magnetic Stirrers |
| Other reagents required | Normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4), Glacial acetic acid (analytical reagent, acetic acid concentration ≥99.5%), Absolute ethanol |
| Storage | 2-8℃ |
| Valid period | 12 months |
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the formal experiment and the detection range (2.92-40 μmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Human plasma | 1 |
| Rat serum | 1 |
| Rat plasma | 1 |
| Mouse serum | 1 |
| Mouse plasma | 1 |
| 10% Rat heart tissue homogenate | 1 |
| 10% Rat liver tissue homogenate | 1 |
| 10% Rat spleen tissue homogenate | 1 |
| 10% Rat lung tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Rat brain tissue homogenate | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
MDA in the catabolite of lipid peroxide can react with thiobarbituric acid (TBA) and produce red compound, which has a maximum absorption peak at 532 nm.
Kit Components & Storage
| Item | Component | Size 1 (48 T) | Size 2 (96 T) | Storage |
| Reagent 1 | Clarificant | 3 mL × 1 vial | 3 mL × 1 vial | 2-8℃, 12 months |
| Reagent 2 | Acid Reagent | 2 mL × 1 vial | 4 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent |
Powder × 1 vial | Powder × 1 vial | 2-8℃, 12 months shading light |
| Reagent 4 | 50 μmol/L Standard | 5 mL × 1 vial | 5 mL × 1 vial | 2-8℃, 12 months |
| Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces | |||
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 4.60 | 25.30 | 32.50 |
| %CV | 4.4 | 4.0 | 3.9 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 4.60 | 25.30 | 32.50 |
| %CV | 8.0 | 6.9 | 6.7 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 97.8%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (μmol/L) | 12 | 18 | 29 |
| Observed Conc. (μmol/L) | 11.6 | 17.8 | 28.3 |
| Recovery rate (%) | 96.9 | 99 | 97.5 |
Sensitivity
The analytical sensitivity of the assay is 1.13 μmol/L MDA This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve:
As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (μmol/L) | 0 | 5 | 10 | 10 | 20 | 25 | 30 | 40 |
| Average OD | 0.041 | 0.071 | 0.093 | 0.122 | 0.155 | 0.182 | 0.21 | 0.269 |
| Absoluted OD | 0 | 0.03 | 0.052 | 0.081 | 0.114 | 0.142 | 0.17 | 0.229 |