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Malondialdehyde (MDA) Fluorometric Assay Kit
SKU: E-BC-F007-96
Malondialdehyde (MDA) Fluorometric Assay Kit
| SKU # | E-BC-F007 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=520 nm/550 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | MDA |
| Sample type | Serum, plasma, animal tissue, cell |
| Sensitivity | 0.04 μmol/L |
| Detection Range | 0.04-10 μmol/L |
| Detection Method | Fluorescence method |
| Precision | Inter-assay CV: 4.5-7.0% | intra-assay CV: 2.3-4.3% |
| Storage | This product can be stored at 2~8°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| Human serum | 2-4 |
| Human plasma | 1 |
| Rat serum | 1 |
| Rat plasma | 1 |
| Mouse serum | 2-4 |
| Mouse plasma | 1 |
| 10% Mouse brain tissue homogenate | 1 |
| 10% Rat brain tissue homogenate | 1 |
| 10% Mouse liver tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 2-4 |
| 9.2×10^6 CHO cells | 1 |
The diluent of serum (plasma) or animal tissue is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4); The diluent of cells is CAMT lysis buffer. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
MDA in the catabolite of lipid peroxide can react with thiobarbituric acid (TBA) and produce red compound, which is proportional to the fluorescence intensity at the excitation wavelength of 520 nm and emission wavelength of 550 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Clarificant | 6 mL ×1 vial | 12 mL × 1 vial | 2-8°C, 12 months |
| Reagent 2 | Acid Reagent | 2 mL ×1 vial | 4 mL × 1 vial | 2-8°C, 12 months |
| Reagent 3 | TBA Reagent | Powder ×1 vial | Powder × 1 vial | 2-8°C, 12 months shading light |
| Reagent 4 | 20 μmol/L Standard | 5 mL ×1 vial | 5 mL × 1 vial | 2-8°C, 12 months |
| Reagent 5 | CAMT Lysis Buffer | 20 mL ×1 vial | 40 mL × 1 vial | 2-8°C, 12 months |
| Black 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) | 0.80 | 4.50 | 7.30 |
| %CV | 4.3 | 3.3 | 2.3 |
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) | 0.80 | 4.50 | 7.30 |
| %CV | 4.5 | 7.0 | 5.6 |
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 97%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc.(μmol/L) | 0.7 | 3.5 | 7.5 |
| Observed Conc.(μmol/L) | 0.7 | 3.5 | 7.3 |
| Recovery Rate (%) | 95 | 99 | 97 |
Sensitivity
The analytical sensitivity of the assay is 0.04 μmol/L. 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 fluorescence 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 | 0.5 | 1 | 2 | 4 | 6 | 8 | 10 |
| Fluorescence value | 57 | 164 | 262 | 486 | 893 | 1339 | 1780 | 2105 |
| 62 | 168 | 258 | 495 | 924 | 1373 | 1757 | 2083 | |
| Average fluorescence value | 59 | 166 | 260 | 490 | 909 | 1356 | 1768 | 2094 |
| Absoluted fluorescence value | 0 | 107 | 1 | 431 | 850 | 1297 | 1709 | 2035 |