计算溶液所需的质量、体积或浓度。
土壤碱性磷酸酶 (S-AKP/ALP) 活性检测试剂盒 (pNPP, 微量法)
有货
为什么选择此级别
BioReagent,标曲法,比色法,适用于分析 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。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
产品介绍
土壤磷酸酶是一类催化土壤有机磷矿化的酶，其活性的高低直接影响着土壤中有机磷的分解转化及其生物有效性，是评价土壤磷素生物转化方向与强度的指标。根据其最适pH 土壤磷酸酶通常分为酸性、中性和碱性三种类型。
本试剂盒主要检测的是土壤中碱性磷酸酶活性。其原理为土壤中的碱性磷酸酶 (S-AKP/ALP)能够在碱性环境中催化磷酸对硝基苯酯 (PNPP) 生成黄色对硝基苯酚 (PNP)，该产物在405nm 处有最大吸收峰，通过测定PNP的生成量即可表征土壤碱性磷酸酶的活性。
S1526065 | 组件 | 物料外观 | 48T | 96 T | 储存条件 | 单次测试使用量 |
S1526065A | 检测缓冲液 | 液体 | 15 mL | 30 mL | 2-8℃. | 按需 |
S1526065B | 显色底物 | 粉末 | 10 mg | 20 mg | 2-8℃.避光. | 按需 |
S1526065C | 终止显色液 | 液体 | 10 mL | 20 mL | 2-8℃. | 200 µL |
S1526065D | 标准品 (10 mM) | 液体 | 1.0 mL | 1.5 mL | 2-8℃.避光. | 按需 |
注意：该试剂盒的各组分均有一定刺激性或毒性，请规范穿戴实验服和手套，避免接触到皮肤。
使用方法
1. 自备试剂耗材
(1) 96 孔板，1.5 mL离心管以及30-50 目筛
(2) 37℃恒温培养箱或者水浴锅，酶标仪或可见分光光度计（能测 405 nm 处的吸光度）
(3) 甲苯，去离子水。
2. 样品检测
建议正式实验前，选取2个样本做预测定，了解实验样品情况，熟悉流程，避免样本和试剂浪费。
(1) 样品制备：取新鲜土样自然风干或 37℃烘箱风干，研磨后过30-50目筛备用。
(2) 显色底物工作液配制：用检测缓冲液将显色底物溶解，终浓度为1 mg/mL。
(3) 标曲稀释：取100 μL标准品(10 mM) + 400 μL去离子水，配制成2 mM的标准品工作液，然后将标准品工作液用去离子水分别稀释成0.1 mM，0.2 mM，0.4 mM，0.8 mM，1.2 mM，1.6 mM和2.0 mM，对应的摩尔质量分别为：5 nmol，10 nmol，20 nmol，40 nmol，60 nmol，80 nmol和100 nmol。
(4) 上机检测：在1.5 mL离心管中按照下表，依次加入各试剂
试剂名称 | 测定管 | 对照管 | 空白管 | 标准管 |
土样 | 0.02 g | 0.02 g | - | - |
甲苯 | 50 μL | 50 μL | 50 μL | - |
显色底物工作液 | 200 μL | - | 200 μL | 200 μL |
检测缓冲液 | - | 200 μL | - | - |
标准品 | - | - | - | 50 μL |
振荡混匀后于37 ℃避光孵育1 h。 | ||||
终止反应液 | 200 μL | 200 μL | 200 μL | 200 μL |
混匀后，12000 rpm离心10 min，吸取200 μL上清至96孔板中，读取A₄₀₅。ΔA=A测定 - A对照；Δ标=A标准 - A空白 |
3. 结果计算
(1) 标准曲线：Y=0.0264*X + 0.001(仅供参考)；X是PNP的摩尔质量(nmol)，Y是△标准。
(2) 酶活性定义：每克土壤每小时水解底物（PNPP）产生 1nmol产物（PNP）为一个酶活单位，计为U。
U=[(△A-0.001)÷0.0264]÷W÷T×D
W-土壤样品质量，g。
T-反应时间，1 h。
D-稀释倍数，未稀释即为1。
注意事项
1. 如果样品信号值过低，可加大土壤样品用量；反之，减少土壤样品用量。
2. 该试剂盒各组分均具有一定刺激性或者毒性，在操作中，请规范穿戴实验服和手套，避免接触到皮肤。
3. 该试剂盒仅做科研用途。
Product Introduction
Soil phosphatases are a group of enzymes that catalyze the mineralization of soil organic phosphorus. Their activity levels directly govern the decomposition, transformation and bioavailability of organic phosphorus in soil, serving as an indicator for evaluating the direction and intensity of biological phosphorus transformation in soil. Based on their optimum pH values, soil phosphatases are generally categorized into three types: acid phosphatase, neutral phosphatase and alkaline phosphatase.
This assay kit is designed primarily for detecting alkaline phosphatase activity in soil. The detection principle is as follows: soil alkaline phosphatase (S-AKP/ALP) catalyzes p-nitrophenyl phosphate (PNPP) to produce yellow p-nitrophenol (PNP) under alkaline conditions. This product has an absorption maximum at 405 nm, and the activity of soil alkaline phosphatase can be characterized by measuring the production amount of PNP.
S1526065 | Components | Appearance | 48T | 96T | Storage | volume per test |
S1526065A | Assay Buffer | Liquid | 15 mL | 30 mL | 2-8℃. | As need |
S1526065B | Chromogenic substrate | Powder | 10 mg | 20 mg | 2-8℃.Store in the dark. | As need |
S1526065C | Stop Reagent | Liquid | 10 mL | 20 mL | 2-8℃. | 200 µL |
S1526065D | Standard (10 mM) | Liquid | 1.0 mL | 1.5 mL | 2-8℃.Store in the dark. | As need |
Note: All components of this kit may be irritant or toxic. Wear lab coat and gloves appropriately to avoid skin contact.
Usage Method
1. Self-supplied Reagents and Consumables
(1) 96-well plates, 1.5 mL centrifuge tubes, and 30–50 mesh sieves.
(2) Incubator or water bath set at 37°C; microplate reader or visible spectrophotometer (capable of measuring absorbance at 405 nm).
(3) Toluene, deionized water.
2. Sample Assay
It is recommended to perform a pre-test with 2 samples prior to the formal experiment to characterize sample properties and get familiar with the operating procedure, so as to avoid waste of samples and reagents.
(1) Sample Preparation: Air-dry fresh soil samples naturally or dry them in an oven at 37 °C. Grind the dried soil and pass it through a 30-50 mesh sieve for later use.
(2) Preparation of chromogenic substrate working solution: Dissolve the chromogenic substrate in assay buffer to a final concentration of 1 mg/mL.
(3) Standard Curve Dilution: Pipette 100 μL of standard stock solution (10 mM) and mix with 400 μL deionized water to prepare 2 mM standard working solution. Subsequently, dilute the standard working solution with deionized water to obtain gradient concentrations of 0.1 mM, 0.2 mM, 0.4 mM, 0.8 mM, 1.2 mM, 1.6 mM and 2.0 mM, corresponding to molar amounts of 5 nmol, 10 nmol, 20 nmol, 40 nmol, 60 nmol, 80 nmol and 100 nmol, respectively.
(4) Microplate Assay Setup: Add each reagent sequentially into 1.5 mL centrifuge tubes according to the table below.
Reagent Name | Sample Tube | Control Tube | Blank Tube | Standard Tube |
Soil sample | 0.02 g | 0.02 g | - | - |
Toluene | 50 μL | 50 μL | 50 μL | - |
Chromogenic substrate working solution | 200 μL | - | 200 μL | 200 μL |
Assay Buffer | - | 200 μL | - | - |
Standard | - | - | - | 50 μL |
Mix thoroughly by shaking, then incubate at 37 °C in dark for 1 h. | ||||
Stop Reagent | 200 μL | 200 μL | 200 μL | 200 μL |
Mix well , centrifuge at 12000 rpm for 10 min. Transfer 200 μL of supernatant to a 96-well plate and record absorbance at 405 nm ( A₄₀₅)。ΔA =ASample - AControl；ΔStandard=AStandard - Ablank |
3. Result Calculation
(1) Standard curve: Y = 0.0264*X + 0.001 (for reference only); X refers to the molar amount of PNP (nmol), and Y represents Δstandard.
(2) Definition of Enzyme Activity: One unit of enzyme activity (U) is defined as the amount of alkaline phosphatase that hydrolyzes the substrate (PNPP) to produce 1 nmol of the product (PNP) per gram of soil per hour.
U = [ (ΔA - 0.001)/0.0264]/W/T×D
W: Mass of soil sample, g
T: Reaction time, 1 h
D: Dilution factor; set as 1 if no dilution is performed
Precautions
1. If the sample signal value is too low, increase the amount of soil sample; conversely, reduce the amount of soil sample.
2. All components of this kit are irritant or toxic. Wear a lab coat and gloves properly during operation to avoid skin contact.
3. This kit is for research use only.
规格
规格或纯度
BioReagent, 比色法, 适用于分析, 标曲法
英文名称
Soil Alkaline Phosphatase (S-AKP/ALP) Activity Assay Kit (pNPP, Microplate Method)
应用
细胞代谢
检测范围
10 U-100 U
检测方法
吸光度法
定量&半定量
定量
检测仪器
酶标仪
检测波长
405 nm
敏感性
Light-sensitive
检测时长
1 h
样本体积
50 μL
储存与运输
储存条件
2-8°C储存
运输条件
低温运输
稳定性与储存
各组分在相应的储存条件下保质期1年。
组分与储存
S1526065 | 组件 | 物理外观 | 48T | 96T | 储存 | 单次测试使用量 |
S1526065A | 检测缓冲液 | 液体 | 10 mL | 20 mL | 2-8℃. | 按需 |
S1526065B | 显色底物 | 粉末 | 10 mg | 20 mg | 2-8℃.避光 | 按需 |
S1526065C | 终止显色液 | 液体 | 10 mL | 20 mL | 2-8℃. | 200 µL |
S1526065D | 标准品 (10 mM) | 液体 | 1.0 mL | 1.5 mL | 2-8℃.避光 | 按需 |
技术文档
高级数据
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C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
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