Soil Catalase(S-CAT) Activity Assay Kit/SLBC0100
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Product Detail
Soil Catalase(S-CAT) Activity Assay Kit
Note: Take two or three different samples for prediction before test.
Operation Equipment: Spectrophotometer
Catalog Number: SLBC0100
Size:50T/24S
Components:
Reagent I: Liquid 0.5 mL×1. Storage at 4℃. Before use, take 0.05 mL of Reagent I and add 9.95 mL of distilled water to dilute it for use or prepare it in proportion. The left reagent stored at 4℃.
Reagent Ⅱ: Powder ×1. Storage at 4℃. Add 2 mL of distilled water before using to dissolve it. The left reagent should be stored at 4℃.
Reagent Ⅲ: Liquid 6 mL×1. Storage at 4℃.
Product Description:
Soil catalase (S-CAT) is an important enzyme of soil microbial metabolism, which plays an important role in the removal system of H2O2.
Since the absorbance at 240 nm is proportional to the amount of H2O2, the activity of S-CAT can be quantified by measuring the decrease in the absorbance of the reaction solution at 240 nm.
Reagents and Equipment Required but Not Provided.
Table centrifuge, transferpettor, spectrophotometer, water bath, 1 mL quartz cuvette, ice and distilled water.
Procedure:
I. Sample processing:
Fresh soil samples are naturally air-dried or oven to dry at 37℃, then sieved by 30 ~ 50 mesh sieve.
II. Determination procedure:
1. Preheat spectrophotometer for 30 minutes, adjust wavelength to 240 nm and set zero with distilled water.
2. Add reagents with the following list:
Reagent | Test Tube (T) | No Substrate Tube (NSu) | No Soil Tube (NSo) |
Air-dried soil sample (g) | 0.1 | 0.1 | - |
Reagent I (μL) | 1000 | - | 1000 |
Distilled water (μL) | - | 1000 | - |
Shake and culture at 25℃ for 20 minutes. | |||
Reagent Ⅱ (μL) | 25 | 25 | 25 |
Mix thoroughly, centrifuge at 8000 ×g for 5 minutes at room temperature and take all the supernatant. | |||
Reagent Ⅲ (μL) | 120 | 120 | 120 |
Mix thoroughly, detect the absorbance of each tube at 240 nm and noted as AT, ANSU, and ANSO.
Note: Each test tube should be provided with a no substrate tube, and the no soil tube only need test once or twice.
III. Calculation
Unit definition: One unit of enzyme activity is defined as the amount of enzyme catalyzes the degradation of 1 mmol of H2O2 in the reaction system per day at 25℃ every gram of dry soil sample.
S-CAT (U/g) = [(A NSo -A T +A NSu) ×Vra÷(ε×d)×103]÷W÷T=18.9× (A NSo-A T+A NSu)
Vra: Total volume of the reaction system, 1.145×10-3 L;
ε: Molar extinction coefficient of hydrogen peroxide, 43.6 L/mol/cm;
d: Cuvette aperture, 1 cm;
T: Reaction time, 20 minutes=1/72 day;
W: Sample mass, 0.1 g.
Note:
If the absorbed supernatant is still partly turbid, centrifuge it again after adding Reagent Ⅲ.
Recent Product citations:
[1] Hou Q, Wang W, Yang Y, et al. Rhizosphere microbial diversity and community dynamics during potato cultivation[J]. European Journal of Soil Biology, 2020, 98: 103176.