Maltase Activity Assay Kit
SKU: E-BC-K041-M-96
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Maltase Activity Assay Kit
| SKU # | E-BC-K041-M |
| Detection Instrument | Microplate reader (500 nm-520 nm, optimum wavelength: 505 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Sample Type | Animal tissue |
| Sensitivity | 6.32 U/mL |
| Detection Range | 6.32-750 U/mL |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 60 min |
| Precision | Average inter-assay CV: 5.700% | Average intra-assay CV: 2.700% |
| Other instruments required | Micropipettor, Vortex mixer, Centrifuge, Water bath, Incubator |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (6.32-750 U/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Rat heart tissue homogenate | 1 |
| 10% Rat liver tissue homogenate | 1 |
| 10% Rat spleen tissue homogenate | 1 |
| 10% Mouse intestinal tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| 10% Mouse brain tissue homogenate | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
Maltase catalyze the corresponding substrate to produce monosaccharide. Monosaccharide produce hydrogen peroxide under the action of oxidase. Hydrogen peroxide react with chromogenic agent to form red product. The activity of maltase can be calculated by detection the optical density with spectrophotometer at 505 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Substrate | Powder × 1 vial | 2-8℃, 12 months |
| Reagent 2 | Buffer Solution | 8 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Stop Solution | 3.5 mL × 1 vial | 2-8℃, 12 months |
| Reagent 4 | Chromogenic Agent A | 13 mL × 1 vial | 2-8℃, 12 months shading light |
| Reagent 5 | Chromogenic Agent B | 13 mL × 1 vial | 2-8℃, 12 months shading light |
| Reagent 6 | 50 mmol/L Glucose Standard Solution | 1 mL × 1 vial | 2-8℃, 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 mouse intestinal 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 (U/mL) | 16.50 | 247.00 | 584.00 |
| %CV | 3.3 | 2.6 | 2.2 |
Inter-assay Precision
Three mouse intestinal tissue were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/mL) | 16.50 | 247.00 | 584.00 |
| %CV | 6.1 | 5.6 | 5.4 |
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%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 4.5 | 13 | 24 |
| Observed Conc. (mmol/L) | 4.5 | 13.3 | 25.4 |
| Recovery rate (%) | 101 | 102 | 106 |
Sensitivity
The analytical sensitivity of the assay is 6.32 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 (mmol/L) | 0 | 2.5 | 5 | 10 | 15 | 20 | 25 | 30 |
| Average OD | 0.154 | 0.236 | 0.322 | 0.496 | 0.672 | 0.829 | 1.036 | 1.188 |
| Absoluted OD | 0 | 0.082 | 0.168 | 0.342 | 0.518 | 0.675 | 0.882 | 1.034 |