Mitochondrial Complex IV (Cytochrome C Oxidase ) Activity Assay Kit
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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.
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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 | |||
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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.
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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 |
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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 |
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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.