Solid-Cellulase(S-CL) Activity Assay Kit
Spectrophotometry
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ชุดทดสอบ Solid-Cellulase (S-CL) Activity Assay Kit ใช้กันอย่างแพร่หลายในงานวิจัยและการประยุกต์ใช้ในอุตสาหกรรมเพื่อวัดกิจกรรมของเอนไซม์เซลลูเลส ซึ่งมีความสำคัญในการย่อยสลายเซลลูโลสให้เป็นน้ำตาลโมเลกุลเล็ก ในอุตสาหกรรมเชื้อเพลิงชีวภาพ ชุดทดสอบช่วยเพิ่มประสิทธิภาพของเอนไซม์เซลลูเลสในการแปลงชีวมวลพืชให้เป็นน้ำตาลที่หมักได้สำหรับการผลิตเอทานอล ในภาคสิ่งทอ ชุดทดสอบใช้ในการประเมินประสิทธิภาพของเซลลูเลสในกระบวนการขัดเงาชีวภาพและการซักแบบหิน เพื่อให้เนื้อผ้านุ่มและได้พื้นผิวตามต้องการ นอกจากนี้ ยังใช้ในอุตสาหกรรมกระดาษและเยื่อกระดาษเพื่อประเมินประสิทธิภาพของเซลลูเลสในการปรับปรุงคุณภาพกระดาษและกระบวนการรีไซเคิล ในงานวิจัยด้านการเกษตร ชุดทดสอบช่วยในการศึกษากิจกรรมของเซลลูเลสในจุลินทรีย์ในดิน ซึ่งมีส่วนช่วยในการทำความเข้าใจการย่อยสลายสารอินทรีย์และสุขภาพของดินให้ดียิ่งขึ้น การประยุกต์ใช้ยังขยายไปถึงการแปรรูปอาหาร ซึ่งช่วยในการปรับปรุงเนื้อสัมผัสและคุณภาพของผลิตภัณฑ์ เช่น น้ำผลไม้และสินเบเกอรี่ โดยการย่อยสลายเส้นใยเซลลูโลส
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Soil Cellulase (S-CL) Activity Assay Kit
Product code: 67126
Method: Spectrophotometric method
Package size: 50T
Product Introduction
Soil cellulase (S-CL) is mainly derived from soil microorganisms. S-CL catalyzes crop straw cellulose to produce glucose, an important carbon-source nutrient.
This kit uses the 3,5-dinitrosalicylic acid method to determine S-CL activity by measuring the reducing sugars produced during catalyzed cellulose degradation.
Sample example: lawn soil. Actual readings may vary depending on testing conditions and instruments.
Package List and Storage
| Item Code | Component | Specification | Storage |
|---|---|---|---|
| 67126.1 | Reagent I | Self-provided | / |
| 67126.2 | Reagent II | 15 mL | 2–8°C |
| 67126.3 | Reagent III | 60 mL | 2–8°C |
| 67126.4 | Reagent IV | 10 mL | 2–8°C, protected from light |
| 67126.5 | Standard | 10 mg | 2–8°C |
| 67126.m | Manual | 1 copy | / |
Quality and Safety Information
| Raw Material or Packaging Name | Quality Standard | Main Toxicity |
|---|---|---|
| Reagent I | —— | —— |
| Reagent II | —— | —— |
| Reagent III | —— | —— |
| Reagent IV | —— | —— |
| Standard | —— | —— |
Transport and Storage
Transport: This product is transported with ice packs.
Storage: Store according to the instructions. Shelf life is 180 days.
Instructions for Use
1. Sample Processing
Air-dry fresh soil samples naturally or oven-dry at 37°C. Pass the dried samples through a 30–50 mesh sieve.
2. Reagent Preparation
- Reagent I: Toluene, 7 mL × 1 bottle, stored at 4°C. This reagent is self-prepared.
- Standard: 10 mg anhydrous glucose. Before use, add 1 mL distilled water to dissolve and prepare a 10 mg/mL glucose solution. Store at 2–8°C for up to two weeks. For longer storage, dissolve with saturated benzoic acid solution.
3. Assay Procedure
- Preheat the visible spectrophotometer or microplate reader for 30 min. Set the wavelength to 540 nm and zero with distilled water.
- Dilute the 10 mg/mL standard solution with distilled water to prepare 1, 0.8, 0.6, 0.4, and 0.2 mg/mL standard solutions for testing.
Standard Solution Dilution
| No. | Concentration Before Dilution (mg/mL) | Standard Solution Volume (µL) | Distilled Water Volume (µL) | Concentration After Dilution (mg/mL) |
|---|---|---|---|---|
| 1 | 10 | 100 | 900 | 1 |
| 2 | 1 | 160 | 40 | 0.8 |
| 3 | 1 | 120 | 80 | 0.6 |
| 4 | 1 | 80 | 120 | 0.4 |
| 5 | 1 | 40 | 160 | 0.2 |
In the following experiments, each standard tube requires 50 µL standard solution. Do not directly measure absorbance at this dilution step.
Assay Operation
| Component or Step | Control Tube | Assay Tube | Standard Tube | Blank Tube |
|---|---|---|---|---|
| Air-dried soil sample | 0.25 g | 0.25 g | - | - |
| Reagent I | 125 µL | 125 µL | - | - |
| Control tube treatment | Boil for 15 min. Wrap with sealing film to prevent the cap from popping off. | - | - | - |
| Assay tube treatment | - | Shake to mix thoroughly and place at room temperature for 15 min. | - | - |
| Reagent II | 250 µL | 250 µL | - | - |
| Reagent III | 1000 µL | 1000 µL | - | - |
| Distilled water | 250 µL | 250 µL | - | - |
| Saccharification | Shake and mix well. Incubate in a 40°C water bath for 1 h, then boil for 15 min. Wrap with sealing film to prevent the cap from popping off. Cool, centrifuge at 10000 rpm at room temperature for 10 min, and collect the supernatant as the saccharified solution. | Shake and mix well. Incubate in a 40°C water bath for 1 h, then boil for 15 min. Wrap with sealing film to prevent the cap from popping off. Cool, centrifuge at 10000 rpm at room temperature for 10 min, and collect the supernatant as the saccharified solution. | - | - |
| Saccharified solution | 50 µL | 50 µL | - | - |
| Standard | - | - | 50 µL | - |
| Distilled water | - | - | - | 50 µL |
| Reagent IV | 150 µL | 150 µL | 150 µL | 150 µL |
| Color development | Mix well and boil in a boiling water bath for 15 min. Wrap with sealing film to prevent the cap from popping off. Cool. | Mix well and boil in a boiling water bath for 15 min. Wrap with sealing film to prevent the cap from popping off. Cool. | Mix well and boil in a boiling water bath for 15 min. Wrap with sealing film to prevent the cap from popping off. Cool. | Mix well and boil in a boiling water bath for 15 min. Wrap with sealing film to prevent the cap from popping off. Cool. |
| Distilled water | 1050 µL | 1050 µL | 1050 µL | 1050 µL |
Mix well after cooling. Transfer 1000 µL into a 1 mL glass cuvette and measure absorbance at 540 nm. Record the values as Acontrol, Ameasured, Astandard, and Ablank.
Calculate ΔAmeasured= Ameasured- Acontroland ΔAstandard= Astandard- Ablank.
For each assay tube, set up one control tube. Blank tubes and the standard curve only need to be run 1–2 times.
Activity Calculation
1. Standard Curve
Use the standard tube concentration X (mg/mL) and absorbance ΔAstandard(Y) to establish a standard curve. Substitute ΔAmeasuredinto the standard curve to calculate the sample concentration X (mg/mL).
2. S-CL Enzyme Activity
Unit definition: The amount of enzyme in each g soil sample that produces 1 mg glucose per day is defined as one enzyme activity unit.
S-CL enzyme activity (U/g soil sample) = X × Vtotal reaction÷ W ÷ T = 156 × X
- X: Sample concentration calculated from the standard curve, mg/mL
- T: Reaction time, 1 h = 1/24 d
- Vtotal reaction: Total volume of the reaction system, 1.625 mL
- W: Sample mass, 0.25 g
Precautions
- This 50T kit can test 24 samples. Before formal measurement, it is recommended to select 2–3 samples with large expected differences for a preliminary test.
- Instruments and supplies required but not provided: visible spectrophotometer, water bath or metal bath, adjustable pipette, 1 mL glass cuvette, 30–50 mesh sieve, toluene, and distilled water.
- If the absorbance of the sample assay tube is too low, 0.01, the reaction time can be extended by increasing the 40°C water-bath saccharification time, possibly to 24 h or longer. The formula must be adjusted accordingly during calculation.
- Alternatively, the saccharified solution volume in the color development step can be adjusted while reducing the distilled water volume. In some cases, the distilled water volume may be completely replaced by saccharified solution. The standard curve must be modified accordingly.
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