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| Product Name | Amylase Activity Colorimetric Assay Kit |
| Catalog No. | ETR-HMM-0059 |
| Description | A colorimetric assay kit for the quantitative measurement of alpha-amylase enzyme activity in biological samples using a defined starch or blocked chromogenic substrate. Alpha-amylase cleaves internal alpha-1,4-glycosidic bonds in starch substrates, generating reducing sugars or releasing chromogenic dyes in proportion to enzyme activity. |
| Intended Use | Measurement of amylase activity in serum, plasma, urine, and salivary samples for clinical biomarker research associated with pancreatitis diagnosis, salivary gland function studies, and evaluation of amylase in food and industrial enzyme applications. |
| Principle / Technology | Alpha-amylase catalyzes the endohydrolysis of alpha-1,4-glucosidic linkages in starch polysaccharides to yield oligosaccharides; in this kit, a blocked-oligosaccharide substrate (such as maltotetraose coupled to a chromogenic p-nitroaniline group) is cleaved by amylase to release the p-nitroaniline chromophore, which absorbs at 405 nm; the rate of p-nitroaniline release is directly proportional to amylase activity. |
| Detection Method | Colorimetric kinetic or endpoint absorbance measurement at 405 nm using a microplate spectrophotometer |
| Sample Type | Serum, plasma, urine, saliva, duodenal fluid, tissue homogenates (pancreas, salivary gland), food samples, industrial fermentation products |
| Performance Range / Specifications | Amylase activity range: 0.1–1,000 U/L depending on sample type and dilution; typical serum reference range 30–110 U/L; calibrator with defined activity included; one unit defined as amount producing 1 μmol reducing sugar equivalents per minute per liter at 37°C |
| Sensitivity / LOD | Lower detection limit approximately 0.5 U/L in diluted serum using kinetic measurement over 3 minutes |
| Specificity | The blocked chromogenic substrate selectively measures endo-amylase activity; exoglycosidases (amyloglucosidase, maltase) cannot release the chromophore without endohydrolysis; glucoamylase present in some biological samples is excluded through pH and substrate design optimization |
| Reaction Conditions / Protocol | Dilute sample appropriately in Sample Dilution Buffer (recommended 1:10 for serum); equilibrate reaction mixture (Substrate Buffer with substrate) at 37°C for 5 minutes; add 5–10 μL diluted sample to pre-warmed substrate; measure absorbance at 405 nm kinetically every 30 seconds for 3 minutes; calculate activity using calibrator comparison (U/L = ΔOD sample/min ÷ ΔOD calibrator/min × calibrator activity) |
| Components / Formulation | Chromogenic Substrate Solution (blocked oligosaccharide substrate at optimized concentration in HEPES buffer, pH 7.0), Activator Solution (sodium chloride, calcium chloride, zinc chloride), Amylase Calibrator (human serum-based reference with certified activity in U/L), Sample Dilution Buffer |
| Storage Conditions | Substrate Solution and Activator: 2–8°C protected from light, stable 12 months; Calibrator: -20°C after reconstitution, stable 4 weeks; Solution components stable at 2–8°C for 6 months after first opening |
| Shelf Life | 12 months from date of manufacture |
| Package Specifications | 200 assays, 500 assays, 2,000 assays (96-well format) |
| Product Form | Liquid ready-to-use reagents with human serum-based calibrator |
| Quality Control | Each lot validated against certified reference serum calibrators from international reference laboratories; bias within ±5% of assigned value; CV ≤3% within-run precision; stable reaction at 37°C for 3 minutes kinetic window |
| Key Features | Traceability to international amylase activity calibrators; liquid-stable format without lyophilized reconstitution; direct comparison with clinical automated analyzer methods using the same substrate class; applicable to both kinetic and fixed-time endpoint modes |
For research use only, not for clinical use.
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