Solid-β-Glucosidase(S-β-GC) Activity Assay Kit
enzyme labeling method
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Soil β-Glucosidase (S-β-GC) Activity Assay Kit - Microplate Method
Product Code
67045
Product Introduction
Soil β-glucosidase (S-β-GC) catalyzes the hydrolysis of glycosidic bonds between aryl or hydrocarbon groups and glycosyl groups to produce glucose. It is an important component of the cellulolytic enzyme system and plays important physiological roles in soil microbial carbohydrate metabolism.
S-β-GC catalyzes p-nitrophenyl-β-D-glucopyranoside to produce p-nitrophenol, which has characteristic absorbance at 400 nm.
Package Contents
| Size | Item Code | Component | Quantity |
|---|---|---|---|
| 100T | 67045.1 | Reagent I | 1 bottle |
| 100T | 67045.2 | Reagent II | 1 bottle |
| 100T | 67045.3 | Reagent III | 1 bottle |
| 100T | 67045.m | Instruction Manual | 1 copy |
Toluene must be prepared separately.
Quality Standards and Safety Information
| Raw Material or Packaging Name | Quality Standard | Main Toxicity |
|---|---|---|
| Reagent I | -- | -- |
| Reagent II | -- | -- |
| Reagent III | -- | -- |
Transportation and Storage
| Transportation | Transport with ice packs. |
|---|---|
| Storage | Store Reagent I at -20°C. Store the remaining components at 2-8°C, protected from light. |
| Shelf Life | 180 days. |
Instructions
1. Sample Processing
Air-dry fresh soil samples naturally, or oven-dry at 37°C. Pass the dried samples through a 30-50 mesh sieve.
2. Assay Procedure
- Preheat the microplate reader for at least 30 min and set the wavelength to 400 nm.
- Before use, add 10 mL double-distilled water to Reagent I. Store any unused portion at -20°C.
- Add samples and reagents according to the table below. Set one control tube for each measurement tube.
| Component or Step | Assay Tube | Control Tube |
|---|---|---|
| Air-dried soil sample (g) | 0.02 | 0.02 |
| Toluene (μL) | 10 | 10 |
| Mixing step | Shake and mix well at room temperature for 15 min | Shake and mix well at 90°C for 15 min |
| Reagent I (μL) | 130 | - |
| Distilled water (μL) | - | 130 |
| Reagent II (μL) | 160 | 160 |
Mix well, incubate in a 37°C water bath for 1 h, then immediately boil in a boiling water bath for 5 min. Cap tightly to prevent water loss. Cool under running water.
Centrifuge at 10000g and 25°C for 10 min. Collect the supernatant and add the following reagents to a 96-well plate.
| Component | Assay Tube | Control Tube |
|---|---|---|
| Supernatant (μL) | 70 | 70 |
| Reagent III (μL) | 130 | 130 |
Mix thoroughly and let stand at room temperature for 2 min. Measure the absorbance at 400 nm as A, then calculate ΔA.
ΔA = Ameasurement- Acontrol
3. S-β-GC Activity Calculation
The regression equation determined under standard conditions is:
y = 0.0016x - 0.0027
In this equation, x is the standard concentration (μmol/L), and y is the absorbance value.
Unit definition: The production of 1 μmol p-nitrophenol per day by 1 g soil sample is defined as one enzyme activity unit.
S-β-GC activity (μmol/d/g) = (ΔA + 0.0027) ÷ 0.0016 × Vtotal reaction÷ W ÷ T = 225 × (ΔA + 0.0027)
- T: reaction time, 1 h = 1/24 d
- Vtotal reaction: total volume of the reaction system, 3 × 10-4L
- W: sample mass, 0.02 g
Precautions
- This 100T kit can test 48 samples.
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