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Thiobarbituric Acid Reactants (TBARS) Fluorometric Assay Kit
SKU: E-BC-K298-F-500
Thiobarbituric Acid Reactants (TBARS) Fluorometric Assay Kit
| SKU # | E-BC-K298-F |
| Detection Instrument | Fluorescent Microplate reader(λex/em = 520 nm/550 nm) |
| Detection method | Fluorescence method |
Product Details
Properties
| Synonyms | TBARS |
| Sample type |
Serum, plasma, animal tissue, cells |
| Sensitivity | 0.09 μmol/L |
| Detection range | 0.09-10 μmol/L |
| Detection Method | Fluorescence method |
| Assay type | Quantitative |
| Assay time | 80 min |
| Precision | Average inter-assay CV: 2.800% | Average intra-assay CV: 1.700% |
| Other instruments required | Vortex mixer, Water bath, Micropipettor |
| Other reagents required | Acetic acid, Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| 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 (0.09-10 μmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Rat serum | 1 |
| 10% Mouse brain tissue homogenization | 1 |
| Human plasma | 1 |
| Mouse serum | 1 |
| 10% Rat liver tissue homogenization | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
TBARS and TBA can react under high temperature and acid conditions and then form a pink compound, the concentration of which is linearly related to the concentration of TBARS in the sample. The TBARS concentration can be calculated by measuring the fluorescence values at the excitation wavelength of 530 nm and the emission wavelength of 550 nm.
Kit Components & Storage
| Item | Component | Size 1 (48 T) | Size 2 (96 T) | Storage |
| Reagent 1 | Clarificant | 12 mL × 1 vial | 12 mL × 1 vial | 2-8°C, 12 months |
| Reagent 2 | Acid Reagent | 6 mL × 1 vial | 12 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℃, 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) | 1.20 | 4.60 | 7.30 |
| %CV | 1.8 | 1.8 | 1.5 |
Inter-assay Precision
Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 1.20 | 4.60 | 7.30 |
| %CV | 2.5 | 2.6 | 3.3 |
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 96%.
| Parameters | Standard 1 | Standard 2 | Standard 3 |
| Expected Conc. (μmol/L) | 0.85 | 3.6 | 7.8 |
| Observed Conc. (μmol/L) | 0.8 | 3.5 | 7.6 |
| Recovery rate (%) | 94.2 | 96 | 96.9 |
Sensitivity
The analytical sensitivity of the assay is 0.09 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 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 | 0.5 | 1 | 2 | 4 | 6 | 8 | 10 |
| Average fluorescence value | 50.2 | 87.5 | 122.7 | 190.1 | 310.0 | 458.5 | 584.4 | 702.8 |
|
Absoluted fluorescence value |
0 | 37.3 | 72.5 | 140.0 | 260.0 | 408.3 | 534.3 | 652.6 |