Aconitase(ACO) Activity Assay Kit
Aconitase(ACO) Activity Assay Kit
| SKU # | E-BC-K656-M |
| Detection Instrument | Microplate reader (230-250 nm, optimum wavelength: 240 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | ACO |
| Sample type | Animal tissue |
| Sensitivity | 2.32 U/L |
| Detection range | 2.32-60.19 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time |
25 min |
| Precision | Average inter-assay CV: 9.500% | Average intra-assay CV: 5.900% |
| Other instruments required | Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl) , PBS(0.01 M，pH 7.4) |
| Storage | -20℃ |
| 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% Rat liver tissue homogenate | 6-10 |
| 10% Mouse kidney tissue homogenate | 4-6 |
| 10% Mouse liver tissue homogenate | 6-10 |
| 10% Mouse brain tissue homogenate | 4-6 |
| 10% Rat lung tissue homogenate | 6-10 |
| 10% Mouse muscle tissue homogenate | 2-4 |
Note: The diluent is extraction solution B. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Aconitase catalyzes isocitrate to produce cis-aconitase. Cis-aconitase has a characteristic absorption peak at 240 nm. The activity of this enzyme was calculated by measuring the production rate of cis-aconitase.
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 | -20℃, 12 months |
| Reagent 2 | Extraction Solution B | 25 mL × 1 vial | 50 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Protease Inhibitor | 0.7 mL × 1 vial | 1.4 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 4 | Buffer Solution | 15 mL × 1 vial | 30 mL × 1 vial | -20℃, 12 months |
| Reagent 5 | Substrate | 0.15 mL × 2 vials | 0.15 mL × 4 vials | -20℃, 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 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 (U/L) | 7.50 | 22.60 | 48.50 |
| %CV | 10.0 | 9.4 | 9.1 |
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 (U/L) | 7.50 | 22.60 | 48.50 |
| %CV | 5.3 | 5.6 | 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 103%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 11.2 | 28.5 | 53 |
| Observed Conc. (U/L) | 11.2 | 27.9 | 56.7 |
| Recovery rate (%) | 104 | 98 | 107 |
Sensitivity
The analytical sensitivity of the assay is 2.32 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.