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for analysis of the inhibition activity of a component in foods
α-Glucosidase Inhibitory Activity Assay Kit

α-Glucosidase deconstructs disaccharides into glucose, and increases the blood glucose level. Therefore, it is possible to prevent increasing the blood glucose level by inhibiting the activity of α-Glucosidase.
Today, many α-Glucosidase inhibitors are discovered in some plants, and vigorously investigated to apply to foods as a suppressor of the blood glucose level.
We released a kit to measure the activity of inhibition of α-Glucosidase. This kit can analyze the inhibition activity of a component in foods by a combination of LabAssay™ Glucose (Sold separately).

1. Link

2. Kit Components

Reagent name

Pkg. size

Number of the vial

The principal ingredient

Enzyme

1 mL

2

α-Glucosidase

Buffer

For 50 mL

2

Maleic Acid/Disodium Maleate

Substrate 1

For 20 mL

2

D-(+)-Maltose

Substrate 2

For 20 mL

2

Sucrose

Positive control

100 mg

1

Acarbose

 

  • Other supplies required
    ●LabAssay™ Glucose
    ●96 wells transparent microplate
    ●Plate reader (with an absorption filter at 505nm)
    ●Micro-pipettes and pipette tips
    ●Incubator (capable of keeping at 37 deg.)

3. Principle of the Assay

α-Glucosidase Inhibitory Activity Assay Kit

4.Assay Procedure

1. Reagent Preparation

α-Glucosidase Inhibitory Activity Assay Kit

Preliminary test for determination of the dilution rate
  Maltose Sucrose

Substrate solution

Maltose solution 50 μL

Sucrose solution 50μL

Purified water

25 μL

Warm for 3 minutes at 37 deg.

Enzyme

Diluted enzyme solution 25 μL

(Example: 5, 10, 20 fold diluent)

Diluted enzyme solution 25 μL

(Example: 1(neat), 2.5, 5 fold dilution)

Mix, and keep accurately for 30 minutes at 37 deg.

Purified water

400 μL

Heat immediately in boiling water for 3 minutes, then keep in ice


【Quantification of glucose】
1. Aliquot 100 uL of enzyme reaction solution and diluted glucose standard solution (Concentration: 15, 10, 5, 2.5 mg/dL) into 96 wells microtiter plate, add 150 uL of chromogen reagent (included LabAssay™ Glucose), then mix thoroughly.
2. Warm for 10 minutes at 37 degrees, and measure the absorbance at 505 nm.
3. Determine the dilution rate as the amount of produced glucose becomes the following concentrations.
  For maltose: 10 mg/dL
  For sucrose: 5 mg/dL

Note:
1) The package insert written in Japanese is attached.
  If you need the package insert in English, please contact us.
2) For the determination method of glucose, please refer the package insert of LabAssay™ Glucose.

 

2. Assay Procedure (Enzyme Rection)

Sample Preparation

  Maltose Sucrose

Substrate solution

Maltose solution 50 μL

Sucrose solution 50μL

Sample or Positive control solution
(Purified water as the negative control)

25 μL

Warm for 3 minutes at 37 deg.

 

Diluted enzyme solution 25 μL

(At the dilution rate determined by the preliminary test)

Diluted enzyme solution 25 μL

(At the dilution rate determined by the preliminary test)

Mix, and keep accurately for 30 minutes at 37 deg.

Purified water

400 μL

Heat immediately in boiling water for 3 minutes, then keep in ice

 

Blank Preparation

 

Maltose

Sucrose

Substrate solution

Maltose solution 50 μL

Sucrose solution 50μL

Sample solution or Purified water

25 μL

Purified water

400 μL

Diluted enzyme solution

Diluted enzyme solution 25 μL

(At the dilution rate determined by the preliminary test)

Diluted enzyme solution 25 μL

(At the dilution rate determined by the preliminary test)

Heat immediately in boiling water for 3 minutes, then keep in ice

 

3. Glucose quantification

Perform the same method using LabAssay™ Glucose as the preliminary test.

 

4. IC50 Calculation

A) Set the yield of glucose of control as 100% α-Glucosidase activity.
  Calculate the specific activity of each concentration of samples.

B) Plot the concentration of sample/positive control at X-axis of semi logarithmic paper and specific activity of α-Glucosidase activity at Y-axis.
  The concentration should be the final concentration in Enzyme reaction solution.
  Calculate the concentration at the point of the specific activity is 50% (IC50) from approximate curve.

α-Glucosidase Inhibitory Activity Assay Kit

5. Product List

α-Glucosidase Inhibitory Activity Assay Kit

Product Name Pkg. Size Grade Wako Cat. No. Storage Condition
α-Glucosidase Inhibitory Activity Assay Kit 96 tests for Food Analysis

297-78601

Keep at -20℃

Glucose quantification kit

Product Name Pkg. Size Grade Wako Cat. No. Storage Condition
LabAssay™ Glucose 1,000 tests for Cellbiology

298-65701

Keep at 2-10℃

α-Glucosidase inhibitors

Product Name Pkg. Size Grade Wako Cat. No. Storage Condition
Salacinol Standard, Synthetic 5 mg for Food Analysis

194-17841

Keep at -20℃
1-Deoxynojirimycin 10 mg for Biochemistry

043-24931

Keep at 2-10℃
N-Butyl-1-deoxynojirimycin 10 mg

020-12631

Keep at -20℃
Voglibose 10 mg for Pharmacology Research

229-01891

Keep at RT
Miglitol 10 mg

138-16221

Keep at -20℃
100 mg

134-16223

Keep at -20℃
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