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Mitochondrial Complex II Activity Assay Kit
SKU: E-BC-K150-M-96
Mitochondrial Complex II Activity Assay Kit
| SKU # | E-BC-K150-M |
| Detection Instrument | Microplate reader (590-610 nm, optimum wavelength: 600 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Sample type | Animal tissue, cell |
| Sensitivity | 0.54 U/L |
| Detection Range | 0.54-17.66 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 15 min |
| Precision | Average inter-assay CV: 5% | Average intra-assay CV: 4% |
| Other instruments required | Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl), PBS(0.01 M，pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Detection Principle
Coenzyme Q, the catalytic product of mitochondrial complex II, can further reduce 2, 6-dichloroindoxol, which has a characteristic absorption peak at 600 nm. Therefore, the activity of mitochondrial complex II can be quantified by measure the change OD value at 600 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 vials | 2-8°C, 12 months |
| Reagent 2 | Extraction Solution B | 15 mL × 1 vial | 30 mL × 1 vial | 2-8°C, 12 months |
| Reagent 3 | Inhibitor | 0.8 mL × 1 vial | 0.8 mL × 2 vials | 2-8°C, 12 months, shading light |
| Reagent 4 | Buffer Solution | 10 mL×1 vial | 20 mL×1 vial | 2-8°C, 12 months |
| Reagent 5 | Substrate A | 0.8 mL×1 vial | 1.6 mL×1 vial | 2-8°C, 12 months, shading light |
| Reagent 6 | Substrate B | 1.5 mL×1 vial | 1.5 mL×2 vials | 2-8°C, 12 months, shading light |
| Reagent 7 | Substrate C | 0.6 mL×1 vial | 1.2 mL×1 vial | 2-8°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 rat 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 (U/L) | 2.35 | 12.60 | 15.50 |
| %CV | 4.3 | 3.8 | 3.9 |
Inter-assay Precision
Three rat 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 (U/L) | 2.35 | 12.60 | 15.50 |
| %CV | 4.9 | 5.2 | 4.9 |
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%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (μmol/L) | 4.5 | 9.8 | 14.5 |
| Observed Conc. (μmol/L) | 4.5 | 9.6 | 13.3 |
| Recovery rate (%) | 101 | 98 | 92 |
Sensitivity
The analytical sensitivity of the assay is 0.54 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.