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The β-Amylase Activity Assay Kit is a spectrophotometric assay designed for the quantitative determination of β-amylase (EC 3.2.1.2) activity in plant or biological samples. β-Amylase catalyzes the hydrolysis of α-1,4-glycosidic linkages at the non-reducing ends of starch to release maltose and other reducing sugars. This kit is based on the colorimetric measurement of reducing sugars, which reduce 3,5-dinitrosalicylic acid to produce a red-brown compound detectable at 540 nm.
The kit is ideal for comparative studies, enzyme kinetics, and screening experiments in plant physiology, agricultural science, and biochemical research.
Key Features
- ✅ Sensitive and Quantitative: Detects β-amylase activity via reducing sugar production using a colorimetric method.
- ✅ Distinction Between α- and β-Amylase: By heat-inactivating α-amylase or β-amylase selectively, the kit allows for specific quantification of each enzyme's activity.
- ✅ High Reproducibility: Standard curve-based analysis ensures consistency across replicates.
- ✅ Comprehensive Components: Includes all necessary reagents and standards for 50 tests.
Kit Components
| Component | Specification | Storage |
|---|---|---|
| Reagent I | 65 mL × 1 bottle | 4°C |
| Reagent II | Powder × 1 vial | 4°C (Reconstitute with 35 mL dH₂O) |
| Standard | 10 mg glucose powder | 4°C (Reconstitute to 10 mg/mL) |
Required Equipment (User Supplied)
- Spectrophotometer (540 nm)
- Water baths (70°C and 40°C)
- Centrifuge (6000 ×g)
- Mortar/homogenizer
- Transfer pipettes
- 1 mL cuvettes
- Distilled water
Procedure Summary
-
Sample Extraction
Homogenize 0.1 g sample in 0.8 mL water, extract at room temp (15 min), centrifuge, and dilute to 10 mL for amylase stock solution. Further dilute 1:5 for total amylase measurement. -
Standard Curve Preparation
Dilute 10 mg/mL glucose solution to prepare six standards (0.00625–0.2 mg/mL). -
Reaction Setup
Perform tests with α-amylase, total amylase, and standards using a combination of heat treatments and reagent additions. Incubate, cool, and read absorbance at 540 nm. -
Calculation
- Generate a standard curve (Abs vs. glucose concentration)
- Calculate α- and total amylase activity
- Derive β-amylase activity by subtracting α from total activity
- Express results as U/g fresh weight or U/mg protein
Calculation Formulas
- α-Amylase (U/g) = (2 × x₁) ÷ W
- Total Amylase (U/g) = (10 × x₂) ÷ W
-
β-Amylase (U/g) = [(10 × x₂) − (2 × x₁)] ÷ W
Where: - x₁, x₂ = glucose concentration from standard curve
- W = sample weight in grams
- Cpr = protein concentration (if using protein normalization)
Storage & Stability
- Store all components at 4°C upon receipt.
- Reconstituted reagents should be used promptly or stored as specified.
References
- Dziedzoave NT et al., Food Control, 2010: “Influence of variety and growth environment on β-amylase activity of flour from sweet potato.”
No resources are currently available for this product.
The β-Amylase Activity Assay Kit is a spectrophotometric assay designed for the quantitative determination of β-amylase (EC 3.2.1.2) activity in plant or biological samples. β-Amylase catalyzes the hydrolysis of α-1,4-glycosidic linkages at the non-reducing ends of starch to release maltose and other reducing sugars. This kit is based on the colorimetric measurement of reducing sugars, which reduce 3,5-dinitrosalicylic acid to produce a red-brown compound detectable at 540 nm.
The kit is ideal for comparative studies, enzyme kinetics, and screening experiments in plant physiology, agricultural science, and biochemical research.
Key Features
- ✅ Sensitive and Quantitative: Detects β-amylase activity via reducing sugar production using a colorimetric method.
- ✅ Distinction Between α- and β-Amylase: By heat-inactivating α-amylase or β-amylase selectively, the kit allows for specific quantification of each enzyme's activity.
- ✅ High Reproducibility: Standard curve-based analysis ensures consistency across replicates.
- ✅ Comprehensive Components: Includes all necessary reagents and standards for 50 tests.
Kit Components
| Component | Specification | Storage |
|---|---|---|
| Reagent I | 65 mL × 1 bottle | 4°C |
| Reagent II | Powder × 1 vial | 4°C (Reconstitute with 35 mL dH₂O) |
| Standard | 10 mg glucose powder | 4°C (Reconstitute to 10 mg/mL) |
Required Equipment (User Supplied)
- Spectrophotometer (540 nm)
- Water baths (70°C and 40°C)
- Centrifuge (6000 ×g)
- Mortar/homogenizer
- Transfer pipettes
- 1 mL cuvettes
- Distilled water
Procedure Summary
-
Sample Extraction
Homogenize 0.1 g sample in 0.8 mL water, extract at room temp (15 min), centrifuge, and dilute to 10 mL for amylase stock solution. Further dilute 1:5 for total amylase measurement. -
Standard Curve Preparation
Dilute 10 mg/mL glucose solution to prepare six standards (0.00625–0.2 mg/mL). -
Reaction Setup
Perform tests with α-amylase, total amylase, and standards using a combination of heat treatments and reagent additions. Incubate, cool, and read absorbance at 540 nm. -
Calculation
- Generate a standard curve (Abs vs. glucose concentration)
- Calculate α- and total amylase activity
- Derive β-amylase activity by subtracting α from total activity
- Express results as U/g fresh weight or U/mg protein
Calculation Formulas
- α-Amylase (U/g) = (2 × x₁) ÷ W
- Total Amylase (U/g) = (10 × x₂) ÷ W
-
β-Amylase (U/g) = [(10 × x₂) − (2 × x₁)] ÷ W
Where: - x₁, x₂ = glucose concentration from standard curve
- W = sample weight in grams
- Cpr = protein concentration (if using protein normalization)
Storage & Stability
- Store all components at 4°C upon receipt.
- Reconstituted reagents should be used promptly or stored as specified.
References
- Dziedzoave NT et al., Food Control, 2010: “Influence of variety and growth environment on β-amylase activity of flour from sweet potato.”
No resources are currently available for this product.
Specifications
| Pack Size | For 50 Assays, For 100 Assays |