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Malondialdehyde (MDA) Colorimetric Assay Kit (Cell Samples)
SKU: E-BC-K028-M-500
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Malondialdehyde (MDA) Colorimetric Assay Kit (Cell Samples)
| SKU # | E-BC-K028-M |
| Detection Instrument | Microplate reader (530-540 nm, optimum wavelength: 532 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | MDA |
| Sample Type | Cells |
| Sensitivity | 0.29 nmol/mL |
| Detection Range | 0.29-100 nmol/mL |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 65 min |
| Precision | Average inter-assay CV: 3.500% | Average intra-assay CV: 3.300% |
| Other instruments required | Micropipettor, Centrifuge, Incubator, Water bath, Vortex mixer |
| Other reagents required | 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 pre-experiment and the detection range (0.29-100 nmol/mL).
| Sample type | Dilution factor |
| A549 cells | 1 |
| HepG2 cells | 1 |
| 293T cells | 1 |
Note: The diluent is reagent 5.
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 (500 Assays) | Storage |
| Reagent 1 | Clarificant | 30 mL × 1 vial | 2-8℃, 12 months |
| Reagent 2 | Acid Reagent | 30 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent | 50 mL × 3 vials | 2-8℃, 12 months shading light |
| Reagent 4 | 10 nmol/mL Standard | 5 mL × 1 vial | 2-8℃, 12 months |
| Reagent 5 | Extracting Solution | 30 mL × 1 vial | 2-8℃, 12 months |
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 HepG2 cell 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 (nmol/mL) | 2.60 | 34.50 | 88.50 |
| %CV | 3.6 | 3.2 | 3.1 |
Inter-assay Precision
Three HepG2 cell samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (nmol/mL) | 2.60 | 34.50 | 88.50 |
| %CV | 3.1 | 3.8 | 3.6 |
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 95%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (nmol/mL) | 12 | 49 | 91 |
| Observed Conc. (nmol/mL) | 11.2 | 48.0 | 85.5 |
| Recovery rate (%) | 93 | 98 | 94 |
Sensitivity
The analytical sensitivity of the assay is 0.29 nmol/mL 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.