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Mitochondrial Complex Ⅴ (F0F1-ATPase/ATP Synthase) Activity Assay Kit
SKU: E-BC-K153-M-96
Mitochondrial Complex Ⅴ (F0F1-ATPase/ATP Synthase) Activity Assay Kit
| SKU # | E-BC-K153-M |
| Detection Instrument | Microplate reader (330-350 nm, optimum wavelength: 340 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | ATP Synthase |
| Sample Type | Animal tissue, cell |
| Sensitivity | 12.13 U/L |
| Detection Range | 12.13-74.21 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 15 min |
| Precision | Average inter-assay CV: 6.500% | Average intra-assay CV: 5.600% |
| Other instruments required | Centrifuge, 37℃ incubator |
| Storage | -20℃ |
| Valid period | 12 months |
Detection Principle
Mitochondrial complex V is also known as F0F1-ATP synthase. ATP is hydrolyzed by F0F1-ATP synthase to produce ADP, and ADP converts NADH into oxidized coenzyme I (NAD) after enzyme conversion reaction. Therefore, the activity of mitochondrial complex IV can be quantified by measure the change OD value at 340 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extraction Solution A |
50 mL ×1 vial | 50 mL × 2 vial | -20°C, 12 months |
| Reagent 2 | Extraction Solution B |
25 mL × 1 vial | 50 mL × 1 vial | -20°C, 12 months |
| Reagent 3 | Protease Inhibitor |
0.8 mL ×1 vial | 0.8 mL × 2 vials | -20°C, 12 months, shading light |
| Reagent 4 | Buffer Solution | 15 mL × 1 vial | 30 mL × 1 vial | -20°C, 12 months |
| Reagent 5 | Substrate A | Liquid ×1 vial | Liquid × 2 vials | -20°C, 12 months, shading light |
| Reagent 6 | Substrate B | Powder ×1 vial | Powder × 2 vials | -20°C, 12 months, shading light |
| Reagent 7 | Substrate C | Powder ×1 vial | Powder × 2 vials | -20°C, 12 months, shading light |
| Reagent 8 | Substrate D | Powder ×1 vial | Powder × 2 vials | -20°C, 12 months, shading light |
| Reagent 9 | Inhibitor | 0.05 mL × 1 vial | 0.1 mL × 1 vial | -20°C, 12 months, shading light |
| UV 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 kidney 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 (U/L) | 21.50 | 38.50 | 65.60 |
| %CV | 5.8 | 5.6 | 5.4 |
Inter-assay Precision
Three mouse kidney tissue samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/L) | 21.50 | 38.50 | 65.60 |
| %CV | 6.4 | 6.3 | 6.8 |
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 101%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 25 | 32.4 | 70.2 |
| Observed Conc. (U/L) | 25.8 | 32.1 | 70.9 |
| Recovery rate (%) | 103 | 99 | 101 |
Sensitivity
The analytical sensitivity of the assay is 12.13 U/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.