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β-Amylase Activity Assay Kit
SKU: E-BC-K005-M-500
β-Amylase Activity Assay Kit
| SKU # | E-BC-K005-M |
| Detection Instrument | Microplate reader(535 nm-540 nm, optimum wavelength: 540 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Sample Type | Plant tissue |
| Sensitivity | 0.97 U/g tissue |
| Detection Range | 0.97-34.74 U/g tissue |
| Detection Method | Colorimetric method |
| Assay Time | 40 min |
| Precision | Inter-assay CV: 3.2% | intra-assay CV: 2.3% |
| 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 |
|---|---|
| 1% Epipremnum aureum tissue homogenate | 1 |
| 1% Green pepper tissue homogenate | 1 |
| 1% Corn grain tissue homogenate | 1 |
| 1% Daucus carota tissue homogenate | 1 |
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. β-amylase was inactivated by the property of amylase not to be heat-resistant, and then the enzyme activity of total amylase and α-amylase is determined. So the activity of β-amylase can be calculated indirectly.
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 epipremnum aureum 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 (mg/mL) |
1.50 | 15.00 | 25.00 |
| %CV | 2.6 | 2.0 | 2.3 |
Inter-assay Precision
Three epipremnum aureum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
|
Mean (mg/mL) |
1.50 | 15.00 | 25.00 |
| %CV | 3.0 | 3.4 | 3.2 |
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 96%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mg/mL) | 0.3 | 0.7 | 1.1 |
| Observed Conc. (mg/mL) | 0.3 | 0.7 | 1.0 |
| Recovery rate (%) | 98 | 95 | 95 |
Sensitivity
The analytical sensitivity of the assay is 0.97 U/g tissue. 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 | 0.156 | 0.308 | 0.497 | 0.700 | 0.886 | 1.013 | 1.238 | 1.440 |
| 0.155 | 0.313 | 0.494 | 0.704 | 0.851 | 1.003 | 1.236 | 1.411 | |
| Average OD | 0.156 | 0.310 | 0.496 | 0.702 | 0.869 | 1.008 | 1.237 | 1.426 |
| Absoluted OD | 0 | 0.154 | 0.496 | 0.546 | 0.713 | 0.852 | 1.081 | 1.270 |