Acetyl-CoA Colorimetric Assay Kit
Acetyl-CoA Colorimetric Assay Kit
| SKU # | E-BC-K652-M |
| Detection Instrument | Microplate reader (330-350 nm, optimum wavelength: 340 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | Acetyl-CoA |
| Sample type | Animal tissue |
| Sensitivity | 150 nmol/mL |
| Detection range | 150-500 nmol/mL |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time |
15 min |
| Precision | Average inter-assay CV: 5.300% | Average intra-assay CV: 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 lung tissue homogenate | 1 |
| 10% Rat liver tissue homogenate | 1 |
| 10% Rat spleen tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Mouse lung tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
Note: The diluent is extracting solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Malate and NAD+ form oxoacetate and NADH under the action of malate dehydrogenase, then acetyl-coa and oxoacetate form coenzyme A and citrate under the action of citrate synthetase. The content of acetyl-coA can be calculated by measuring the change of absorbance value at 340 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extracting Solution | 60 mL × 1 vial | 60 mL × 2 vials | -20℃, 12 months |
| Reagent 2 | Buffer Solution | 20 mL × 1 vial | 40 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Enzyme Reagent A | Liquid ×1 vial | Liquid ×2 vial | -20℃, 12 months, shading light |
| Reagent 4 | Enzyme Reagent B | Power × 1 vial | Power × 2 vials | -20℃, 12 months, shading light |
| Reagent 5 | Substrate | Power × 1 vial | Power × 2 vials | -20℃, 12 months, shading light |
| Reagent 6 | 500 nmol/mL Standard | 0.16 mL×1 vial | 0.32 mL×1 vial | -20℃, 12 months, shading light |
| 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 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 (nmol/mL) | 225.00 | 348.00 | 405.00 |
| %CV | 4.4 | 4.0 | 3.6 |
Inter-assay Precision
Three rat 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 (nmol/mL) | 225.00 | 348.00 | 405.00 |
| %CV | 5.0 | 5.2 | 5.7 |
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. (nmol/mL) | 185 | 279 | 452 |
| Observed Conc. (nmol/mL) | 192.4 | 287.4 | 461.0 |
| Recovery rate (%) | 104 | 103 | 102 |
Sensitivity
The analytical sensitivity of the assay is 150 nmol/mL. 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.