Soil Neutral Protease Activity Assay Kit
Micro method
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Soil Neutral Protease Activity Assay Kit - Micro Method
Product Information
| Product Code | 67034 |
|---|---|
| Abbreviation | S-NPT |
| Format | 100T |
Product Introduction
Soil protease participates in the conversion of amino acids, proteins, and other nitrogen-containing organic compounds in soil. Its hydrolysis products are one source of nitrogen for higher plants.
S-NPT catalyzes protein hydrolysis in a neutral environment and is related to soil organic matter content, nitrogen, and other soil properties. Under neutral conditions, S-NPT hydrolyzes casein to produce tyrosine. Under alkaline conditions, tyrosine reduces phosphomolybdic acid compounds to form tungsten blue, which has a characteristic absorption peak at 680 nm.
Actual readings may vary under different test conditions and with different instruments.
Product Packing List
| Catalog No. | Component | Quantity |
|---|---|---|
| 67034.1 | Reagent I | 1 bottle |
| 67034.2 | Reagent II | 1 bottle |
| 67034.3 | Reagent III | 1 bottle |
| 67034.4 | Reagent IV | 1 bottle |
| 67034.5 | Reagent V | 1 bottle |
| 67034.6 | Reagent VI | 1 tube |
| 67034.7 | Reagent VII | 1 bottle |
| 67034.m | Instruction Manual | 1 copy |
Quality Standards and Safety Information
| Raw Material or Packaging Name | Quality Standard | Main Toxicity |
|---|---|---|
| Reagent I | -- | -- |
| Reagent II | -- | -- |
| Reagent III | -- | -- |
| Reagent IV | -- | -- |
| Reagent V | -- | -- |
| Reagent VI | -- | -- |
| Reagent VII | -- | -- |
Transportation and Storage
| Transportation | Transport with ice packs. |
|---|---|
| Storage | Store at 2-8°C. Shelf life: 180 days. |
Instructions for Use
1. Sample Processing
Naturally air-dry fresh soil samples, or air-dry them in an oven at 37°C. Pass the dried samples through a 30-50 mesh sieve.
2. Reagent Preparation
- Before use, add 6 mL distilled water to Reagent II and dissolve thoroughly. Store unused reagent at 4°C.
- Before use, add 2 mL Reagent VII to Reagent III and heat in a boiling water bath to dissolve. Then add 8 mL Reagent I, mix thoroughly, and set aside for use. Store unused reagent at 4°C.
- Before use, add 30 mL distilled water to Reagent IV and dissolve thoroughly for later use. Store unused reagent at 4°C.
3. Assay Procedure
- Preheat the spectrophotometer or microplate reader for at least 30 min. Set the wavelength to 680 nm and zero with distilled water.
- Add samples and reagents according to the table below.
| Component | Assay Tube | Control Tube |
|---|---|---|
| Air-dried soil sample | 0.02 g | 0.02 g |
| Reagent I | 150 µL | |
| Reagent III | 150 µL |
Mix well and incubate in a 40°C water bath for 30 min. Shake 5-6 times during incubation to allow the soil sample to fully contact the reaction solution.
| Component | Assay Tube | Control Tube |
|---|---|---|
| Reagent II | 50 µL | 50 µL |
Mix well, then centrifuge at 8000 g and 25°C for 10 min. Take the supernatant and add the following reagents to EP tubes.
| Component | Assay Tube | Control Tube | Standard Tube |
|---|---|---|---|
| Supernatant | 60 µL | 60 µL | |
| Reagent VI | 60 µL | ||
| Reagent IV | 280 µL | 280 µL | 280 µL |
| Reagent V | 60 µL | 60 µL | 60 µL |
Mix well and incubate in a 40°C water bath for 20 min. Centrifuge at 8000 g and 25°C for 10 min. Collect 200 µL supernatant and read the absorbance value of each tube at 680 nm, recorded as A.
The standard tube only needs to be measured once. Set one control tube for each assay tube.
S-NPT Activity Calculation
Unit definition: The production of 1 mg tyrosine per g soil sample per day is defined as one S-NPT activity unit.
S-NPT (mg/d/g soil sample) = Cstandard× (Aassay- Acontrol) ÷ Astandard× Vtotal reaction÷ W ÷ T = 24 × (Aassay- Acontrol) ÷ Astandard
| Parameter | Value |
|---|---|
| Cstandard | Standard tube concentration, 0.05 mg/mL |
| Vtotal reaction | Total reaction system volume, 0.4 mL |
| T | Reaction time, 30 min = 1/48 d |
| W | Sample mass, 0.02 g |
Precautions
- This 100T product can test 48 samples.
Visual Reference
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