Soil Urease (SUE) Test Kit

soil urease(SUE) Test Box

Product Details:
Measurement significance:

S-UE can hydrolyze urea to produce ammonia and carbonic acid. The urease activity in soil is positively correlated with the number of microorganisms, organic matter content, total nitrogen, and available nitrogen content in the soil. Soil urease activity reflects the nitrogen status of the soil.

Measurement principle:

Determination of urease hydrolysis of urea using indophenol blue colorimetric methodNH3-N。

Equipment and supplies to be provided:

Visible spectrophotometer/ELISA reader, water bath, adjustable pipette, trace quartz colorimetric dish/96 well plate, mortar, ice, toluene (express delivery not allowed), and distilled water.
in the sampleAA extraction:

1. According to the ratio of tissue mass (g): reagent volume (mL) of 1:5-10 (it is recommended to weigh about 0.1g of tissue and add 1mL of reagent one), homogenize at room temperature, then transfer to a 1.5 mL EP tube, cover tightly (to prevent water loss), and place in a boiling water bath for extraction for 15 minutes; After cooling the tap water, centrifuge 8000g at 4 ℃ for 10 minutes, and place the supernatant on ice for testing.

2. Bacteria or cultured cells: First collect bacteria or cells into a centrifuge tube, centrifuge and discard the supernatant; According to the ratio of the number of bacteria or cells (104) to the volume of reagent (mL) of 500-1000:1 (it is recommended to add 1mL of reagent 1 to 5 million bacteria or cells), sonicate the bacteria or cells (ice bath, power of 20% or 200W, sonication for 3 seconds, interval of 10 seconds, repeated 30 times); 8000g， Centrifuge at 4 ℃ for 10 minutes, take the supernatant, and place it on ice for testing.

3. Serum and other liquids: direct detection.

The calculation formula is as follows:

1. Calculation of serum (plasma) LAP activity:

Definition of Unit: EachThe generation of 1 nmol of p-nitroaniline per minute from mL serum (slurry) is defined as one enzyme activity unit.

LAP (nmol/min/mL)=[Δ A × V inverse total ÷ (ε× d) × 109] ÷ V sample ÷ T=205.8 × Δ A

2. Calculation of LAP activity in tissues, bacteria, or cells:

（1) Calculated based on sample protein concentration:

Definition of Unit: EachThe production of 1 nmol of p-nitroaniline per minute by mg tissue protein is defined as one enzyme activity unit.

LAP（nmol/min /mg prot）＝[ΔA×V 反总 ÷（ε×d）×109]÷(V 样 ×Cpr) ÷T=205.8×ΔA÷Cpr

（2) Calculated based on sample fresh weight:

Definition of Unit: EachThe production of 1 nmol of p-nitroaniline per minute by G organization is defined as one enzyme activity unit.

LAP (nmol/min/g fresh weight)=[Δ A × V inverse total ÷ (ε× d) × 109] ÷ (W × V sample ÷ V sample total) ÷ T=205.8 × Δ A ÷ W

（3) Calculated by bacterial or cell density:

Definition of Unit: Each10000 bacteria or cells producing 1 nmol of p-nitroaniline per minute is defined as one enzyme activity unit.

LAP (nmol/min/104 cell)=[Δ A × V inverse total ÷ (ε× d) × 109] ÷ (2000 × V sample ÷ V sample total) ÷ T=0.103 × Δ A

V anti total: total volume of the reaction system, 2 × 10-4 L; ε: molar extinction coefficient of p-nitroaniline, 9.72 × 103 L/mol/cm; d: Colorimetric dish diameter, 1cm; V-sample: sample volume added, 0.05 mL; V-sample total: extraction solution volume added, 1 mL; T: reaction time, 2 min; Cpr: sample protein concentration, mg/mL； W: Sample quality, g; 2000: total number of bacteria or cells, 20 million.

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Notes:

1. Reagents 3, 4, and standard in the reagent kit must be prepared before use and stored away from light. If not fully prepared, they should be stored at 4 ℃ and used within 3 days.

To ensure the accuracy of the experimental results, 1-2 samples need to be taken for preliminary experiments. If the measured absorbance value is too high (higher than 2.5), dilute it with distilled water before measuring.

3. There is no absorption peak at 570nm when reacting with indene ketone, therefore, the measurement results at 570nm do not contain the amounts of these two amino acids.

The detection limit is 100 μ mol/L.