Solid-β-Glucosidase(S-β-GC) Activity Assay Kit
Micro method
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Soil-β-Glucosidase (S-β-GC) Activity Assay Kit - Micro Method
Product Information
| Product Code | 67044 |
|---|---|
| Product Size | 100T |
Product Introduction
S-β-GC catalyzes the hydrolysis of glycosidic bonds between aryl or alkyl groups and glycosyl radicals to produce glucose. It is an important component of the cellulolytic enzyme system.
In soil microorganisms, S-β-GC has important physiological functions in carbohydrate metabolism. S-β-GC catalyzes p-nitrophenyl-β-D-glucopyranoside to produce p-nitrophenol, which has characteristic light absorption at 400 nm.
Package Contents
| Code | Component | Quantity |
|---|---|---|
| 67044.1 | Reagent I | 1 bottle |
| 67044.2 | Reagent II | 1 bottle |
| 67044.3 | Reagent III | 1 bottle |
| 67044.m | Manual | 1 copy |
Toluene must be prepared by the user.
Quality and Safety Information
| Raw Material or Packaging Name | Quality Standard | Main Toxicity |
|---|---|---|
| Reagent I | -- | -- |
| Reagent II | -- | -- |
| Reagent III | -- | -- |
Shipping and Storage
| Item | Condition |
|---|---|
| Shipping | Shipped with ice packs. |
| Storage | Store Reagent I at -20°C. Store the remaining components at 2-8°C. |
| Shelf Life | 180 days. |
Instructions
1. Sample Processing
Air-dry fresh soil samples naturally, or dry them in a 37°C oven. Pass the dried samples through a 30-50 mesh sieve.
2. Assay Procedure
- Preheat the spectrophotometer or 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.
| Component or Step | Assay Tube | Control Tube |
|---|---|---|
| Air-dried soil sample | 0.02 g | 0.02 g |
| Toluene | 10 µL | 10 µL |
| Shake and mix at room temperature | 15 min | -- |
| Shake and mix at 90°C | -- | 15 min |
| Reagent I | 130 µL | -- |
| Distilled water | -- | 130 µL |
| Reagent II | 160 µL | 160 µL |
- 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 10000 g and 25°C for 10 min, then collect the supernatant.
- Add the following reagents in an EP tube or 96-well plate.
| Component | Assay Tube | Control Tube |
|---|---|---|
| Supernatant | 70 µL | 70 µL |
| Reagent III | 130 µL | 130 µL |
Mix thoroughly and let stand at room temperature for 2 min. Measure the absorbance at 400 nm.
Calculate ΔA as follows: ΔA = Ameasurement- Acontrol. Set one control tube for each measurement tube.
S-β-GC Activity Calculation
Micro Quartz Cuvette Measurement
The regression equation determined under standard conditions is:
y = 0.0032x - 0.0027
x is the standard concentration (µmol/L), and y is the absorbance value.
Unit definition: The production of 1 µmol p-nitrophenol per day per g soil sample is defined as one enzyme activity unit.
S-β-GC activity (µmol/d/g) = (ΔA + 0.0027) ÷ 0.0032 × Vtotal reaction÷ W ÷ T = 112.5 × (ΔA + 0.0027)
96-Well Plate Measurement
The regression equation determined under standard conditions is:
y = 0.0016x - 0.0027
x is the standard concentration (µmol/L), and y is the absorbance value.
Unit definition: The production of 1 µmol p-nitrophenol per day per 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)
Calculation Parameters
| Parameter | Value |
|---|---|
| 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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