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5’-Nucleotidase (5'-NT) Activity Assay Kit
SKU: E-BC-K196-M-500
5’-Nucleotidase (5'-NT) Activity Assay Kit
| SKU # | E-BC-K196-M |
| Detection Instrument | Microplate reader (550 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | 5’-NT |
| Sample Type | Serum, plasma, animal tissue |
| Sensitivity | 28.0 U/L |
| Detection Range | 28.0-581 U/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 30 min |
| Precision | Average inter-assay CV: 6% | Average intra-assay CV: 3.300% |
| Other instruments required | Pipettor, Water bath, Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Detection Principle
5'-Nucleotide enzyme (5'-NT, EC 3.1.3.5), full name is 5'-ribonucleotide phosphohydrolase, is a special phosphohydrolase that specifically hydrolyzes 5'-phosphoric acid attached to pentose in 5'-nucleotide. This enzyme is widely distributed in the cell membrane of various tissues of human and animal. Only the 5'-NT released by the tissue cells of the hepatobiliary system may enter the blood. Therefore, the source of serum 5'-NT has certain specificity, and the determination of serum 5'-NT has important value for the diagnosis of hepatobiliary diseases.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Working Solution | 10 mL × 1 vial | 20 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 2 | Chromogenic Agent | 5 mL × 1 vial | 10 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 3 | 0.6 mmol/L Inosine Standard Solution | 2 mL × 1 vial | 2 mL × 1 vial | 2-8°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 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 (U/L) | 52.00 | 227.00 | 406.00 |
| %CV | 3.8 | 3.4 | 2.7 |
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 (U/L) | 52.00 | 227.00 | 406.00 |
| %CV | 6.1 | 6.3 | 5.6 |
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 105%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 0.21 | 0.34 | 0.45 |
| Observed Conc. (mmol/L) | 0.2 | 0.4 | 0.5 |
| Recovery rate (%) | 108 | 103 | 104 |
Sensitivity
The analytical sensitivity of the assay is 28.0 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.
Standard Curve
As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only.
| Concentration (mmol/L) | 0.00 | 0.12 | 0.24 | 0.3 | 0.36 | 0.42 | 0.48 | 0.60 |
| OD value | 0.046 | 0.073 | 0.100 | 0.116 | 0.126 | 0.143 | 0.153 | 0.192 |
| 0.045 | 0.072 | 0.087 | 0.115 | 0.126 | 0.147 | 0.158 | 0.184 | |
| Average OD | 0.046 | 0.073 | 0.093 | 0.115 | 0.126 | 0.145 | 0.155 | 0.188 |
| Absoluted OD | 0.000 | 0.027 | 0.048 | 0.069 | 0.080 | 0.099 | 0.110 | 0.142 |