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α-Amylase and β-Amylase Activity Assay Kit
SKU: E-BC-K006-M-500
α-Amylase and β-Amylase Activity Assay Kit
| SKU # | E-BC-K006-M |
| Detection Instrument | Microplate reader(535 nm-540 nm, optimum wavelength: 540 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Sample Type | Serum, plasma, saliva, tissue |
| Sensitivity | 0.008 U/mL |
| Detection Range | 0.01-0.56 U/mL |
| Detection Method | Colorimetric method |
| Assay Time | 30 min |
| Precision | Inter-assay CV: 3.6% | intra-assay CV: 2.8% |
| Storage | This product can be stored at 2~8°C for 12 months with shading light. |
| 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 plasma | 10-15 |
| Human serum | 10-15 |
| Human saliva | 15-25 |
| Mouse serum | 15-25 |
| Rat serum | 15-25 |
| 1% Corn grain tissue homogenate | 2-5 |
The diluent is double distilled water. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
The reducing sugar reacts with 3,5-dinitrosalicylic acid under heating conditions to produce a brown-red substance. Amylase activity can be calculated by measuring the OD value at 540 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Substrate | 10 mL ×1 vial | 2-8°C, 12 months |
| Reagent 2 | Chromogenic Agent 2 | 20 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 3 | 10 mg/mL Standard | 1.5 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/mL) |
0.08 | 1.40 | 0.25 |
| %CV | 3.2 | 2.4 | 2.8 |
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/mL) |
0.08 | 1.40 | 0.25 |
| %CV | 3.0 | 4.0 | 3.8 |
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 97%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mg/mL) | 0.3 | 0.7 | 1.1 |
| Observed Conc. (mg/mL) | 0.3 | 0.7 | 1.1 |
| Recovery rate (%) | 99 | 95 | 97 |
Sensitivity
The analytical sensitivity of the assay is 0.008 U/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.
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 (mg/mL) | 0 | 0.2 | 0.4 | 0.6 | 0.8 | 1.0 | 1.2 | 1.4 |
| OD value | 0.144 | 0.310 | 0.483 | 0.688 | 0.829 | 1.016 | 1.224 | 1.361 |
| 0.144 | 0.301 | 0.486 | 0.684 | 0.840 | 1.013 | 1.164 | 1.356 | |
| Average OD | 0.144 | 0.305 | 0.484 | 0.686 | 0.834 | 1.015 | 1.194 | 1.359 |
| Absoluted OD | 0.000 | 0.161 | 0.341 | 0.542 | 0.691 | 0.871 | 1.050 | 1.215 |