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Homocysteine (Hcy) Colorimetric Assay Kit
SKU: E-BC-K854-M-96
Homocysteine (Hcy) Colorimetric Assay Kit
| SKU # | E-BC-K854-M |
| Detection Instrument | Microplate reader (330-350 nm, optimum wavelength: 340 nm) |
|
Detection method |
Colorimetric method |
Product Details
Properties
| Synonyms | Hcy |
| Sample type | Serum (plasma) and urine |
| Sensitivity | 1.95 μmol/L |
| Detection range | 1.95-100.0 μmol/L |
| Detection method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 25 min |
| Precision | Average inter-assay CV: 9% | Average intra-assay CV: 1.700% |
| Other instruments required | Centrifuge |
| 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 |
| Human serum | 1 |
| Human urine | 1 |
Note: The diluent is normal saline (0.9% NaCl). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Oxidative Hcy is converted into free Hcy, which can convert NADH to NAD+ through cyclic catalysis of enzymes, resulting in a decrease of OD value at 340 nm. The Hcy content in the sample is calculated by the rate of decrease.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution A | 10 mL × 1 vial | 20 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 2 | Buffer Solution B | 6 mL × 1 vial | 12 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Substrate | Power × 1 vial | Power × 2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Enzyme Reagent | Power × 2 vials | Power × 4 vials | -20℃, 12 months, shading light |
| Reagent 5 | 28 μmol/L Standard | 0.5 mL × 1 vial | 1 mL × 1 vial | -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 human serum 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) | 5.60 | 35.60 | 84.30 |
| %CV | 2.0 | 1.7 | 1.4 |
Inter-assay Precision
Three human serum 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) | 5.60 | 35.60 | 84.30 |
| %CV | 8.8 | 9.2 | 9.0 |
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%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/L) | 12.6 | 42.8 | 92 |
| Observed Conc. (μmol/L) | 13.2 | 44.1 | 95.7 |
| Recovery rate(%) | 105 | 103 | 104 |
Sensitivity
The analytical sensitivity of the assay is 1.95 μmol/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.