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Adipogenesis Fluorometric Assay Kit
SKU: E-BC-F065-96
Adipogenesis Fluorometric Assay Kit
| SKU # | E-BC-F065 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Sample Type | Cell |
| Sensitivity | 1.4 μmol/L |
| Detection Range | 1.4-32 μmol/L |
| Detection Method | Fluorescence method |
| Assay Time | 20 min |
| Precision | Inter-assay CV: 8.4% | intra-assay CV: 4% |
| Storage | This product can be stored at 2~8°C/-20°C for 12 months with shading light. Please refer to the manual for the specific storage condition of the components. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| 10% Porcine heart tissue homogenate | 3-6 |
| 10% Porcine liver tissue homogenate | 8-10 |
| CHO cell (10^6) | 1 |
| 293T cell (10^6) | 1 |
The diluent is extraction solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Triglycerides were converted by enzyme to produce hydrolytic products, which were catalyzed by the enzyme to produce fluorescent substances. The triglyceride accumulation can be calculated by measuring the fluorescence intensity at the excitation wavelength of 535 nm and the emission wavelength of 587 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Enzyme Working Solution | 25 mL ×1 vial | 2-8℃, 12 months shading light |
| Reagent 2 | Extraction Solution | 50 mL ×1 vial | 2-8℃, 12 months |
| Reagent 3 | Probe | 0.5 mL ×1 vial | -20℃, 12 months shading light |
| Reagent 4 | 1 mmol/L Standard | 0.5 mL ×1 vial | 2-8℃, 12 months shading light |
| 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 porcine heart tissue 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) | 2.00 | 15.00 | 25.00 |
| %CV | 4.3 | 3.8 | 3.9 |
Inter-assay Precision
Three porcine heart tissue 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) | 2.00 | 15.00 | 25.00 |
| %CV | 9.7 | 7.9 | 7.6 |
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 98%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/L) | 8 | 20 | 25 |
| Observed Conc. (μmol/L) | 8.0 | 19.6 | 24.0 |
| Recovery rate (%) | 100 | 98 | 96 |
Sensitivity
The analytical sensitivity of the assay is 1.4 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 | 8 | 16 | 20 | 24 | 32 |
| Fluorescence value | 2488 | 4332 | 6347 | 7177 | 8215 | 10135 |
| 2395 | 4189 | 6213 | 7226 | 7840 | 9835 | |
| Average fluorescence value | 2441.5 | 4260.5 | 6280 | 7201.5 | 8027.5 | 9985 |
| Absoluted fluorescence value | 0 | 1819 | 3838.5 | 4760 | 5586 | 7543.5 |