Mitochondrial Complex IV (Cytochrome C Oxidase ) Activity Assay Kit
Mitochondrial Complex IV (Cytochrome C Oxidase ) Activity Assay Kit
| SKU # | E-BC-K152-M |
| Detection Instrument | Microplate reader (540-560 nm, optimum wavelength: 550 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | Animal tissue |
| Sensitivity | 3.0 U/L |
| Detection Range | 3.0–88.6 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 10 min |
| Precision | Average inter-assay CV: 9.500% | Average intra-assay CV: 5% |
| Other instruments required | Centrifuge |
| Other reagents required | Anhydrous ethanol, PBS(0.01 M，pH 7.4) |
| Storage | -20℃ |
| Valid period | 12 months |
Detection Principle
Mitochondrial complex IV can catalyze the oxidation of reduced cytochrome C to oxidized cytochrome C, which has an absorption wavelength at 550 nm. Therefore, the activity of mitochondrial complex IV can be quantified by measure the change OD value at 550 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extracting Solution A |
50 mL ×1 vial | 50 mL × 2 vials | -20°C, 12 months |
| Reagent 2 | Extracting Solution B |
25 mL × 1 vial | 50 mL × 1 vial | -20°C, 12 months |
| Reagent 3 | Inhibitor | Powder ×1 vial | Powder × 2 vials | -20°C, 12 months, shading light |
| Reagent 4 | Substrate | Powder ×1 vial | Powder × 2 vials | -20°C, 12 months, shading light |
| Reagent 5 | Stabilizer | Powder ×1 vial | Powder × 2 vials | -20°C, 12 months, shading light |
| Reagent 6 | Buffer Solution | 13 mL×1 vial | 26 mL×1 vial | 20°C, 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 rat brain 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) | 8.50 | 34.60 | 70.50 |
| %CV | 5.1 | 5.0 | 4.9 |
Inter-assay Precision
Three rat brain 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) | 8.50 | 34.60 | 70.50 |
| %CV | 9.8 | 9.1 | 9.6 |
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 104%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (μmol/L) | 18.5 | 42.7 | 65.1 |
| Observed Conc. (μmol/L) | 18.7 | 44.8 | 69.0 |
| Recovery rate (%) | 101 | 105 | 106 |
Sensitivity
The analytical sensitivity of the assay is 3.0 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.