Soil Neutral Xylanase(S-NEX) Activity Assay Kit
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Soil Neutral Xylanase (S-NEX) Activity Assay Kit
Product code: 67048
Method: Microplate method
Product Introduction
Xylanase (EC 3.2.1.8) is mainly produced by microorganisms. It catalyzes the hydrolysis of xylan and is also known as pentosanase or hemicellulase.
Xylanase can decompose raw-material cell walls in the brewing and feed industries, as well as beta-glucan. It reduces material viscosity during brewing, promotes the release of effective substances, reduces non-starch polysaccharides in feed, and supports the absorption and utilization of nutrients.
Neutral xylanase (NEX) is generally isolated from microorganisms with an optimal growth pH of 6-8. In a neutral environment, NEX catalyzes the degradation of xylan into reducing oligosaccharides and monosaccharides. These products react with 3,5-dinitrosalicylic acid under boiling water bath conditions to produce a color reaction with a characteristic absorption peak at 540 nm.
The color intensity of the reaction solution is proportional to the amount of reducing sugar produced by enzymatic hydrolysis. NEX activity can be calculated by measuring the increase in absorbance at 540 nm.
Product Packing List
| Size | Code | Component | Quantity |
|---|---|---|---|
| 100T | 67048.1 | Reagent I | 1 bottle |
| 100T | 67048.2 | Reagent II | 1 bottle |
| 100T | 67048.3 | Buffer | 1 bottle |
| 100T | 67048.m | Instructions | 1 copy |
Quality Standards and Safety Information
| Raw Material or Packaging Name | Quality Standard | Main Toxicity |
|---|---|---|
| Reagent I | -- | -- |
| Reagent II | -- | -- |
| Buffer | -- | -- |
Transportation and Storage
| Transportation | Transport with ice packs. |
|---|---|
| Storage | Store at 2-8°C, protected from light. Shelf life: 180 days. |
Product Instructions
1. Sample Processing
Air-dry fresh soil samples and pass them through a 30-50 mesh sieve.
2. Assay Procedure
- Preheat the microplate reader for 30 min and set the wavelength to 540 nm.
- Prepare the control tube and assay tube according to the table below.
| Component | Control Tube | Assay Tube |
|---|---|---|
| Soil sample (g) | 0.02 | 0.02 |
| Buffer (μL) | 150 | 100 |
| Reagent I (μL) | -- | 50 |
- Mix thoroughly and react with shaking at 50°C for 30 min.
- Immediately place the tubes in a 90°C water bath for 10 min.
- Centrifuge at 8000g and 25°C for 10 min.
- Take 100 μL of supernatant.
- Add 100 μL of Reagent II to both the control tube and assay tube.
- Mix thoroughly and develop color in a 90°C water bath for 5 min.
- Transfer 180 μL to a 96-well plate and measure the absorbance at 540 nm.
Record the absorbance values as AControl Tubeand AAssay Tube.
Calculate ΔA as follows: ΔA = AAssay Tube- AControl Tube.
3. Calculation
Standard curve: y = 0.8452x + 0.0058, R2= 0.9989
Enzyme activity definition: Under the conditions of 50°C and pH 6.0, the amount of enzyme required to decompose xylan in each gram of soil per day to produce 1 μmol reducing sugar is defined as one unit of enzyme activity.
Activity (μmol/d/g soil sample) = (ΔA - 0.0058) ÷ 0.8452 × Vtotal reaction× 103÷ W ÷ T ÷ 150 = 56.8 × (ΔA - 0.0058) ÷ W
| Vtotal reaction | Total reaction volume, 0.15 mL |
|---|---|
| T | Reaction time, 1/48 d |
| 1000 | 1 mmol/L = 103μmol/L |
| 150 | Molecular weight of xylose |
| W | Soil sample weight, g |
Precautions
- Before formal measurement, select 2-3 samples with large expected differences for pretesting.
- Required instruments and supplies to be prepared by the user: balance, room-temperature centrifuge, shaker, constant-temperature water bath, microplate reader, and 96-well plate.
- This 100T kit can test 48 samples.
- Ensure a 30 min shaking reaction so the enzyme and substrate are in full contact.
- Use caution with the 90°C water bath to prevent bursting and avoid altering the reaction system.
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