计算溶液所需的质量、体积或浓度。
土壤淀粉酶 (S-AL) 活性检测试剂盒（DNS, 微量法）
有货
为什么选择此级别
BioReagent,无菌 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存,避光。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
土壤淀粉酶（S-AL）是土壤中催化淀粉水解的酶类总称，主要通过微生物富集，能够以随机作用 方式切断淀粉、寡聚或多聚糖分子内的葡萄糖苷键，生成麦芽糖、低聚糖和葡萄糖等，在土壤中糖类物质的转化过程中起重要作用。
检测原理：淀粉酶水解淀粉产生还原糖，可与3,5 二硝基水杨酸反应生成红棕色物质，在540nm处有特征吸收峰，颜色深浅在一定范围内与还原糖量成正比。
适用样本：土壤
注意：正式检测前，建议选择2-3个预期差异较大的样本进行预实验。
自备仪器和试剂
- 酶标仪或可见分光光度计（能测540nm处的吸光度）
- 96孔板或微量玻璃比色皿、可调节式移液枪及枪头
- 恒温水浴锅、恒温摇床、台式离心机、30-50目筛
- 去离子水、甲苯
操作步骤
1、试剂准备
| 试剂名称 | 试剂准备 | 注意事项 |
| Reagent Ⅰ | 即用型，使用前平衡到室温 | 4℃保存 |
| Working Reagent Ⅱ | 临用前配制，48T加入7.2mL去离子水，96T加入14.4mL去离子水，置于70℃水浴，期间不断搅拌或震荡至溶解。 | 用不完的试剂4℃保存一周 |
| Reagent Ⅲ | 即用型，使用前平衡到室温 | 4℃保存 |
注意：Reagent Ⅲ有一定的刺激性，建议在使用过程中做好个人防护。
2、标准品制备
Standard：临用前配制，加入1mL去离子水配成10 mg/mL的葡萄糖标准液；4℃避光保存2周。按照下表所示，进一步稀释标准品：
| 序号 | 10mg/mL Standard（μL） | 去离子水的体积（μL） | 浓度（mg/mL） |
| Std.1 | 40 | 160 | 2 |
| Std.2 | 20 | 180 | 1 |
| Std.3 | 10 | 190 | 0.5 |
| Std.4 | 5 | 195 | 0.25 |
| Std.5 | 2.5 | 197.5 | 0.125 |
| Std.6 | 1.25 | 198.75 | 0.063 |
| Blank | 0 | 200 | 0 |
注意：每次实验都要做一次标准品检测，制作标曲；稀释后的标准品溶液不稳定，必须在4小时内使用。
3、样本制备
注意：推荐使用新鲜样本，如果不立即进行实验，样本可在-80℃保存1个月。测定时，应控制解冻的温度和时间。室温环境下解冻时，需在4h内完成样品解冻。
新鲜土样自然风干或37℃烘箱风干，过30-50目筛。
4、实验步骤
4.1 酶标仪或可见分光光度计预热30min，调节波长到540nm。可见分光光度计用去离子水调零。
4.2 操作表（下述操作在1.5mL EP管中进行）：
| 试剂 | 对照管（μL） | 测定管（μL） | 标准管（μL） | 空白管（μL） |
| 风干土样（g） | 0.04 | 0.04 | 0 | 0 |
| 甲苯 | 20 | 20 | 0 | 0 |
25℃静置15min。
| Reagent Ⅰ | 100 | 100 | 0 | 0 |
| Working Reagent Ⅱ | 0 | 120 | 0 | 0 |
| 去离子水 | 120 | 0 | 0 | 0 |
混匀，37℃恒温摇床震荡24h，25℃，8,000rpm，离心10min，取上清。
| 样本上清 | 120 | 120 | 0 | 0 |
| Standard | 0 | 0 | 120 | 0 |
| 去离子水 | 0 | 0 | 0 | 120 |
| Reagent Ⅲ | 120 | 120 | 120 | 120 |
4.3 充分混匀，90℃水浴5min，待冷却后，取200μL至微量玻璃比色皿/96孔板中测定540nm吸光值，分别记为A对照、A测定、A标准、A空白，计算ΔA测定=A测定-A对照、ΔA标准=A标准-A空白。
注意：
空白孔和标准孔只需测定1次，每次试验需设置对照组。实验之前建议选择2-3个预期差异大的样本做预实验。
每次实验90℃水浴后的冷却时间保持一致，请于30min内完成测定。
5、结果计算
注意：我们为您提供的计算公式，包括推导过程计算公式和简洁计算公式。两者完全相等。建议以加粗的简洁计算公式为最终计算公式。
5.1 标准曲线的绘制
以标准溶液浓度为x轴，ΔA标准为y轴，绘制标准曲线，得到标准方程，将ΔA测定带入方程得到x(mg/mL)。
5.2 酶活的计算
单位的定义：每天每g土样产生1mg还原糖定义为一个土壤淀粉酶活力。
S-AL (U/g 土样)=x×V反总÷W÷T=6×x
参数说明：
V反总：酶促反应总体积，0.24mL；
W：土样质量，0.04g；
T：反应时间，1d。
6、结果展示
图1. 本试剂盒测定土壤中淀粉酶的活性
注意事项
本产品仅供科学研究使用，不适用于临床诊断。为了您的安全和健康，请穿实验服并戴一次性手套操作。
Soil amylase (S‑AL) refers to the collective term for enzymes in soil that catalyze the hydrolysis of starch. It is primarily accumulated by microorganisms and can randomly cleave glycosidic bonds within starch, oligosaccharide, or polysaccharide molecules, producing maltose, oligosaccharides, glucose, etc., playing a significant role in the transformation of carbohydrate substances in soil.
Detection Principle
Amylase hydrolyzes starch to produce reducing sugars, which react with 3,5‑dinitrosalicylic acid to form a reddish‑brown product with a characteristic absorption peak at 540 nm. Within a certain range, the color intensity is proportional to the amount of reducing sugars.
Applicable Sample: Soil
Note: Before formal testing, it is recommended to perform a preliminary experiment with 2‑3 samples expected to show significant differences.
Reagents, consumables and Equipments not provided
Microplate reader or visible spectrophotometer (capable of measuring absorbance at 540 nm)
96‑well plate or micro‑volume glass cuvette, Adjustable pipettes and tips
Constant temperature water bath, Constant temperature shaker, Tabletop centrifuge, 30‑50 mesh sieve
Deionized water, Toluene
Procedure
1. Reagent Preparation
| Reagent Name | Preparation | Notes |
| Reagent Ⅰ | Ready‑to‑use. Equilibrate to room temperature before use. | Store at 4°C. |
| Working Reagent Ⅱ | Prepare immediately before use. For 48T, add 7.2 mL deionized water; for 96T, add 14.4 mL deionized water. Place in a 70°C water bath and stir or shake continuously until dissolved. | Unused reagent can be stored at 4°C for one week. |
| Reagent Ⅲ | Ready‑to‑use. Equilibrate to room temperature before use. | Store at 4°C. |
Note: Reagent Ⅲ is somewhat irritating. Personal protection is recommended during use.
2. Standard Preparation
Standard: Prepare immediately before use. Add 1 mL deionized water to prepare a 10 mg/mL glucose standard solution. Store at 4°C protected from light for up to two weeks. Further dilute the standard as shown in the table below:
| Tube | 10mg/mL Standard（μL） | Deionized Water Volume (µL) | Concentration (mg/mL) |
| Std.1 | 40 | 160 | 2 |
| Std.2 | 20 | 180 | 1 |
| Std.3 | 10 | 190 | 0.5 |
| Std.4 | 5 | 195 | 0.25 |
| Std.5 | 2.5 | 197.5 | 0.125 |
| Std.6 | 1.25 | 198.75 | 0.063 |
| Blank | 0 | 200 | 0 |
Note: A standard curve must be prepared for each experiment. Diluted standard solutions are unstable and must be used within 4 hours.
3. Sample Preparation
Note: Fresh samples are recommended. If not used immediately, samples can be stored at ‑80°C for one month. Control the thawing temperature and time during measurement. When thawing at room temperature, complete the process within 4 hours.
Air‑dry fresh soil samples naturally or in a 37°C oven, then sieve through a 30‑50 mesh screen.
4. Experimental Steps
4.1 Preheat the microplate reader or visible spectrophotometer for 30 minutes and set the wavelength to 540 nm. Zero the visible spectrophotometer with deionized water.
4.2 Operation Table (perform in 1.5 mL EP tubes):
| Reagent | Control Tube (µL) | Test Tube (µL) | Standard Tube (µL) | Blank Tube (µL) |
| Air‑dried soil (g) | 0.04 | 0.04 | 0 | 0 |
| Toluene | 20 | 20 | 0 | 0 |
Let stand at 25°C for 15 min.
| Reagent Ⅰ | 100 | 100 | 0 | 0 |
| Working Reagent Ⅱ | 0 | 120 | 0 | 0 |
| Deionized Water | 120 | 0 | 0 | 0 |
Mix well, shake in a 37°C constant temperature shaker for 24 hours. Centrifuge at 25°C, 8,000 rpm for 10 minutes. Collect the supernatant.
| Sample Supernatant | 120 | 120 | 0 | 0 |
| Standard | 0 | 0 | 120 | 0 |
| Deionized Water | 0 | 0 | 0 | 120 |
| Reagent Ⅲ | 120 | 120 | 120 | 120 |
4.3 Mix thoroughly, incubate in a 90°C water bath for 5 minutes. After cooling, transfer 200 µL to a micro‑volume glass cuvette / 96‑well plate. Measure the absorbance at 540 nm, recorded as AControl, ATest, AStandard, ABlank. Calculate ΔATest = ATest ‑ AControl and ΔAStandard = AStandard ‑ ABlank.
Note:
The blank and standard tubes only need to be measured once. A control group must be set up for each experiment. It is recommended to perform a preliminary experiment with 2‑3 samples showing expected large differences before formal testing.
Keep the cooling time after the 90°C water bath consistent for each experiment. Complete the measurement within 30 minutes.
5. Calculation of Results
Note: We provide two formula sets: the derived calculation formulas and the simplified formulas. They are completely equivalent. The simplified formulas are recommended as the final calculation formulas.
5.1 Plotting the Standard Curve
Plot the standard curve with the standard solution concentration as the x‑axis and ΔAStandard as the y‑axis. Obtain the standard equation. Substitute ΔATest into the equation to obtain x (mg/mL).
5.2 Calculation of Enzyme Activity
Unit Definition: One soil amylase activity unit (U) is defined as the amount of enzyme that produces 1 mg of reducing sugar per day per gram of soil.
S‑AL (U/g soil) = x × VTotal Reaction ÷ W ÷ T = 6 × x
Parameter Description:
VTotal Reaction: Total volume of the enzymatic reaction, 0.24 mL;
W: Soil sample mass, 0.04 g;
T: Reaction time, 1 day.
6. Example Results
Figure 1. Soil amylase activity determined by this kit
Notes
This product is for scientific research use only and is not intended for clinical diagnosis. For your safety and health, please wear a lab coat and disposable gloves during operation.
规格
别名
土壤淀粉酶活性检测试剂盒 微量法 | 淀粉酶活性检测试剂盒（3,5-二硝基水杨酸法）
英文别名
Micro Soil Amylase(S-AL) Activity Assay Kit
规格或纯度
BioReagent, 无菌
英文名称
Soil Amylase (S-AL) Activity Assay Kit (DNS, Micro Method)
应用
细胞代谢
样品类型
土壤
检测范围
0.063-2 mg/mL
检测方法
比色法
定量&半定量
定量
检测仪器
酶标仪
检测波长
540nm
回收率
80%-120%
敏感性
Light-sensitive
储存与运输
储存条件
2-8°C储存,避光
运输条件
低温运输
稳定性与储存
Store at 2-8℃ long term (12 months). Store in the dark.
组分与储存
| 组分货号 | 组分名称 | 物理外观 | 48T | 96T | 储存 | 单次测试使用量 |
| S1508358A | Reagent I | 无色透明液体 | 12 mL | 24 mL | 2-8℃ | 单次测试需100μL |
| S1508358B | Reagent II | 白色粉末 | 1EA | 1EA | 2-8℃ | 按需 |
| S1508358C | Reagent III | 浅黄色透明液体 | 15.6 mL | 31.2 mL | 2-8℃. Store in the dark. | 单次测试需120μL |
| S1508358D | Standard | 白色粉末 | 1EA | 1EA | 2-8℃. Store in the dark. | 按需 |
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0332496 |
分析证书 | S1508358 |
技术文档和文章
溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
Copyright © 2023至今 上海阿拉丁生化科技股份有限公司 保留所有权利。
危险品化学品经营许可证（带存储）