Soil Catalase (S-CAT) Activity Assay Kit/SLBC0105
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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: Microplate reader/ Spectrophotometer
Catalog Number: SLBC0105
Size:100T/48S
Components:
Reagent I: Liquid 0.3 mL×1.Storage at 4℃. The liquid is placed in the EP tube in the reagent bottle. Before use, take 0.05 mL of Reagent I and add 9.95 mL of distilled water to dilute for use or prepare in proportion. The reagent that can not be used up is stored at 4℃;
Reagent Ⅱ: Powder×1. Storage at 4℃. Add 1 mL of distilled water before using to dissolve it. The left reagent should be stored at 4℃.
Reagent Ⅲ: Liquid 3 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 meeasuring the decrease in the absorbance of the reaction solution at 240 nm.
Reagents and Equipment Required but Not Provided.
Table centrifuge, water-bath, transferpettor, micro quartz cuvette/96 well UV flat-bottom plate, spectrophotometer / microplate reader and distilled water.
Procedure
Fresh soil samples are naturally air-dried or oven to dry at 37℃, then sieved by 30 ~ 50 mesh sieve.
1. Preheat spectrophotometer or microplate reader for 30 minutes, adjust the wavelength to 240 nm, 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) | 0.03 | 0.03 |
|
Reagent I (μL) | 260 |
| 260 |
Double distilled water (μL) |
| 260 |
|
shake and culture at 25℃ for 20 minutes. | |||
Reagent Ⅱ (μL) | 10 | 10 | 10 |
Mix thoroughly, centrifuge at 8000 ×g for 5 minutes at room temperature and take all the supernatant. | |||
Reagent Ⅲ (μL) | 30 | 30 | 30 |
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
A. micro cuvette
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)＝[(ANSo-AT+ANSu)×Vra÷(ε×d)×103]÷W÷T=16.5×(ANSo-AT+ANSu)
Vra: Total volume of the reaction system, 3×10-4 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, 0.03 g.
B. 96 well UV plate
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)＝[(ANSo-AT+ANSu)×V ra÷(ε×d)×103]÷W÷T=27.5×(ANSo－AT+ANSu)
Vra: Total volume of the reaction system, 3×10-4 L;
ε: Molar extinction coefficient of hydrogen peroxide, 43.6 L / mol /cm;
d: Cuvette aperture, 0.6 cm;
T: Reaction time, 20 minutes=1/72 day;
W: Sample mass, 0.03 g.
Note:
If the absorbed supernatant is still partly turbid, centrifuge it again after adding Reagent Ⅲ.
Recent Product citations：
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.
References：
[1] 杨兰芳, 曾巧, 李海波, et al. 紫外分光光度法测定土壤过氧化氢酶活性[J]. 土壤通报, 2011, 42(1):207-210.
[2] Johansson L H, Borg L A H. A spectrophotometric method for determination of catalase activity in small tissue samples[J]. Analytical biochemistry, 1988, 174(1): 331-336.