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ATP Chemiluminescence Assay Kit
SKU: E-BC-F002-96
ATP Chemiluminescence Assay Kit
| SKU # | E-BC-F002 |
| Detection Instrument | Chemiluminescence detector |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | ATP |
| Sample type | Animal tissue, cells |
| Sensitivity | 0.003 μmol/L |
| Detection range | 0.003-5 μmol/L |
| Detection method | Fluorescence method |
| Assay time | 15 min |
| Precision | Inter-assay CV: 6.5% | intra-assay CV: 2.2% |
| Storage | This product can be stored at -20°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 |
|---|---|
| 10% Mouse heart tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| 10% Mouse muscle tissue homogenate | 1 |
| 10% Mouse liver tissue homogenate | 1 |
| 10% Mouse brain tissue homogenate | 1 |
| 10% Mouse lung tissue homogenate | 1 |
The diluent is extracting solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Under the catalyzation of luciferase, ATP react with luciferin and emits fluorescence, and the fluorescence intensity is proportional to the concentration of ATP within a certain range.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extracting Solution | 50 mL ×1 vial | 50 mL ×2 vials | -20℃, 12 months |
| Reagent 2 | 100 μmol/L Standard Solution |
1 mL ×1 vial | 1 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Enzyme Reagent | Power ×1 vial | Power × 2 vials | -20℃, 12 months shading light |
| Reagent 4 | Enzyme Diluent | 7 mL ×1 vial | 14 mL × 1 vial | -20℃, 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 mouse liver 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) | 0.40 | 2.30 | 4.00 |
| %CV | 3.5 | 1.6 | 1.5 |
Inter-assay Precision
Three mouse liver 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) | 0.40 | 2.30 | 4.00 |
| %CV | 7.9 | 6.0 | 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 102%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc.(μmol/L) | 0.8 | 2.3 | 3.7 |
| Observed Conc.(μmol/L) | 0.8 | 2.3 | 3.9 |
| Recovery rate (%) | 101 | 99 | 106 |
Sensitivity
The analytical sensitivity of the assay is 0.003 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 | 2.5 | 3 | 4 | 5 |
| Fluorescence value | 20 | 15066 | 31034 | 60635 | 72975 | 86500 | 115711 | 144427 |
| 26 | 15064 | 31034 | 60631 | 72977 | 86502 | 115709 | 144431 | |
| Average fluorescence value | 23 | 15065 | 31034 | 60633 | 72976 | 86501 | 115710 | 144429 |
| Absoluted fluorescence value | 0 | 15041 | 31010 | 60610 | 72953 | 86478 | 115687 | 144405 |