Soil Leucine Aminopeptidase(S-LAP) Activity Assay Kit
enzyme labeling method
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| Test Parameter | Specification |
|---|---|
| Performance test | PASS |
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Soil Leucine Aminopeptidase (S-LAP) Activity Assay Kit - Microplate Method
Product Information
Product code: 67055
S-LAP is a class of enzymes secreted by soil microorganisms that can hydrolyze peptide chains with N-terminal leucine. Changes in S-LAP activity are closely related to certain pathological states of the body.
S-LAP decomposes L-leucyl-p-nitroanilide to produce p-nitroaniline, which has a maximum absorption peak at 405 nm. S-LAP activity can be calculated by measuring absorbance at 405 nm.
Package Contents
| Size | Code | Item | Quantity |
|---|---|---|---|
| 100T | 67055.1 | Reagent I | 1 bottle |
| 100T | 67055.2 | Reagent II | 1 bottle |
| 100T | 67055.m | Instructions | 1 copy |
Quality Standards and Safety Information
| Raw Material or Packaging Name | Quality Standard | Main Toxicity |
|---|---|---|
| Reagent I | -- | -- |
| Reagent II | -- | -- |
Transportation and Storage
| Transportation | Transport with ice packs. |
|---|---|
| Storage | Store Reagent I at 4 °C. Store Reagent II at -20 °C, protected from light. Shelf life is 180 days. |
Instructions for Use
1. Reagent Preparation
Add 15 mL Reagent I to the Reagent II bottle and dissolve thoroughly. If dissolution is difficult, heat in a 60 °C water bath for about 30 min to promote dissolution.
Incubate in a water bath at 37 °C for mammals or 25 °C for other species for 10 min or longer.
Aliquot unused reagent and store at -20 °C. Do not freeze and thaw repeatedly.
2. Operating Procedure
- Preheat the microplate reader for 30 min or longer, set the wavelength to 405 nm, and zero with distilled water.
- Add samples according to the table below.
| Component | Control Tube | Assay Tube |
|---|---|---|
| Fresh soil sample (g) | 0.05 | 0.05 |
| Reagent I (μL) | 300 | - |
| Reagent II (μL) | - | 300 |
- Mix well and react with shaking at 37 °C for 1 h.
- Centrifuge at 8000g, 4 °C for 10 min.
- Transfer 200 μL of supernatant into a 96-well plate and measure the absorbance at 405 nm as A.
- Calculate ΔA = Aassay- Acontrol. Set one control tube for each assay tube.
3. S-LAP Activity Calculation
Unit definition: the generation of 1 μmol p-nitroaniline per day by 1 g soil sample is defined as one unit of enzyme activity.
S-LAP (μmol/d·g-1, soil sample) = ΔA × Vtotal reaction÷ (ε × d) × 106÷ W ÷ T = 29.6 × ΔA
| Vtotal reaction | Total volume of the reaction system, 3 × 10-4L |
|---|---|
| ε | Molar extinction coefficient of p-nitroaniline, 9.72 × 103L/mol·cm-1 |
| d | Optical path length of the cuvette, 0.5 cm |
| T | Reaction time, 1 h = 1/24 d |
| W | Sample mass, 0.05 g |
Precautions
- This 100T kit can test 48 samples.
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