计算溶液所需的质量、体积或浓度。
植物根系活力检测试剂盒 (萘胺, 比色法)
有货
为什么选择此级别
BioReagent,比色法,生化试剂 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
植物根系是活跃的吸收器官和合成器官，根的生长情况和代谢水平即根系活力直接影响植物地上部的生长和营养状况以及最终产量，是植物生长的重要生理指标之一。植物根系活力(根系脱氢酶活性)可以采用TTC还原、萘胺氧化和甲烯蓝吸附等方法测定。根系能氧化萘胺生成红色的萘胺化合物，根系对萘胺的氧化能力与其呼吸强度有密切联系。
该试剂盒的检测原理是在弱酸性条件下，以萘胺为底物，萘胺与对氨基苯磺酸、亚硝酸盐作用生成稳定的红色偶氮化合物，可以根据根系表面着色深浅，通过酶标仪510 nm检测溶液中未被氧化的萘胺的量，以定量测定根系活力。本产品仅用于科研领域，不适用于临床诊断或其他用途。
产品组分
P1515880 | 组分 | 物理外观 | 60 T | 储存条件 | 单次使用量 |
P1515880A | 试剂一 | 液体 | 20 mL | 2-8℃. | 按需 |
P1515880B | 试剂二 | 液体 | 250 mL | 2-8℃. | 按需 |
P1515880C | 试剂三 | 液体 | 25 mL | 2-8℃.避光保存. | 400 μL |
P1515880D | 试剂四 | 液体 | 25 mL | 2-8℃.避光保存. | 400 μL |
P1515880E | 标准品 | 液体 | 5 mL | 2-8℃.避光保存. | 按需 |
注意：标准品为0.5 mg/mL α-萘胺标准品。
自备仪器和试剂
酶标仪或可见分光光度计、水浴锅/恒温培养箱、可调式移液器、滤纸、玻璃比色皿、蒸馏水。
实验流程
1. 样本处理
样本的制备：将根部组织洗净，去除根上的泥土，轻轻擦干，不要过分挤压破坏根系细胞。称取0.5 g于10 mL EP管中。
2. 测定步骤
2.1. 可见分光光度计/酶标仪预热30 min以上，调节波长至510 nm，并用蒸馏水调零。
2.2. 标准品的稀释：将0.5 mg/mL α-萘胺标准品用试剂一稀释为0.5、0.25、0.125、0.0625、0.03125、0.0156、0.0078、0 mg/mL的标准品。(0 mg/mL记为标准空白管)：
2.3. 标准品稀释液：
序号 | 标准品体积 | 试剂一体积(µL) | 浓度(mg/mL) |
Std.1 | 1 mL of 0.5mg/mL标准品 | - | 0.5 |
Std.2 | 500µL of Std.1 | 500 | 0.25 |
Std.3 | 500µL of Std.2 | 500 | 0.125 |
Std.4 | 500µL of Std.3 | 500 | 0.0625 |
Std.5 | 500µL of Std.4 | 500 | 0.03125 |
Std.6 | 500µL of Std.5 | 500 | 0.0156 |
Std.7 | 500µL of Std.6 | 500 | 0.0078 |
Blank | 0 | 500 | 0(标准空白管) |
注意：下述实验中每个标准管需400 µL标准品(注意不要在此步骤直接检测标准品吸光值)。
2.4. 在10 mLEP管按下表步骤加样：
试剂名称(μL) | 测定管 | 空白管 | 标准管 |
样本 | 0.5 g | - | - |
试剂二 | 5000 | 5000 | - |
37℃避光反应，在反应10 min和3 h 10 min时测定管和空白管需要各吸取400 μL反应液进行下述第二步反应。 第二步反应需要先按下表在新的1.5 mL EP管中加入试剂三和试剂四，再吸取400 μL上述第一步反应液于EP管中。注：需要先加试剂三和试剂四，再加反应液，一定不能颠倒顺序！ | - | ||
试剂三 | 400 | 400 | 400 |
试剂四 | 400 | 400 | 400 |
第一步反应液 | 400 | 400 | - |
标准品 | - | - | 400 |
充分混匀，室温显色20 min，取1 mL于玻璃比色皿中，测定在510 nm处的吸光度，显色后需要在20 min内完成检测。10 min所对应的吸光值记作A1，3 h 10 min所对应的吸光值记作A2。计算ΔA测定 = A1测定 - A2测定 ，ΔA空白 =A1空白 -A2空白 ，ΔA =ΔA测定 -ΔA空白 ，ΔA标 =A标准 - A标准空白管 。(标准曲线和空白管只需做1-2次)。
3. 根系活力计算
3.1. 标准曲线的绘制：根据标准管的浓度(x，mg/mL)和吸光度ΔA标(y，ΔA标)，建立标准曲线。根据标准曲线，将ΔA(y，ΔA)带入公式计算样本浓度(x，mg/mL)。
3.2. 按照样本质量计算： 以氧化的α-萘胺的量来表示根系活力根系活力[μg/(g·h)]=x×V试剂二 ×10³÷(W×T)=1666.67×x÷W 10³：单位换算，1 mg/mL=10³ μg /mL；V试剂二 ：试剂二的体积，5 mL；W：根系质量，g；T：反应时间，3 h。
注意事项
1. 在EP管内加入试剂三、试剂四以及样本的顺序不可改变。
2. 如果ΔA结果不在标准曲线范围内，可以减少样本质量(或者减少第一步反应时间)或者增加样本质量(或者增加第一步反应时间)。
显色后需要在20min内完成检测。
Plant roots are active organs for absorption and synthesis. The growth status and metabolic level of roots-known as root activity-directly affect the growth, nutritional status, and final yield of the above-ground parts of plants, making it a key physiological indicator. Root activity (root dehydrogenase activity) can be measured by methods such as TTC reduction, naphthylamine oxidation, and methylene blue adsorption. Roots can oxidize naphthylamine to produce red naphthylamine compounds, and their oxidizing capacity toward naphthylamine is closely related to their respiratory intensity.
The detection principle of this kit is based on the reaction in a weakly acidic environment, using naphthylamine as the substrate. Naphthylamine reacts with sulfanilic acid and nitrite to form a stable red azo compound. By measuring the amount of unoxidized naphthylamine in the solution-indicated by the coloration intensity on the root surface-root activity can be quantitatively determined using a microplate reader at 510 nm. This product is intended for research use only and is not suitable for clinical diagnosis or other purposes.
Kit Contents
P1515880 | Components | Appearance | 60 T | Storage | Quantity Per Test |
P1515880A | Reagent 1 | Liquid | 20 mL | 2-8℃. | As needed |
P1515880B | Reagent 2 | Liquid | 250 mL | 2-8℃. | As needed |
P1515880C | Reagent 3 | Liquid | 25 mL | 2-8℃. Store in the dark. | 400 μL |
P1515880D | Reagent 4 | Liquid | 25 mL | 2-8℃. Store in the dark. | 400 μL |
P1515880E | Standard | Liquid | 5 mL | 2-8℃. Store in the dark. | As needed |
Note: The standard is 0.5 mg/mL α-naphthylamine standard.
User-Prepared Instruments and Reagents:
1. Microplate reader or visible spectrophotometer, water bath/constant-temperature incubator, adjustable pipettes, filter paper, glass cuvettes, distilled water.
Experimental Procedure
1. Sample Preparation: Wash the root tissue thoroughly to remove soil, gently blot dry, and avoid squeezing or damaging the root cells. Weigh 0.5 g of the sample into a 10 mL EP tube.
2. Assay Procedure
2.1. Preheat the visible spectrophotometer/microplate reader for at least 30 min, set the wavelength to 510 nm, and zero the instrument with distilled water.
2.2. Dilution of the standard: Dilute the 0.5 mg/mL α-naphthylamine standard with Reagent 1 to prepare standards at 0.5, 0.25, 0.125, 0.0625, 0.03125, 0.0156, 0.0078, and 0 mg/mL. (The 0 mg/mL solution is recorded as the standard blank tube.)
2.3. Standard diluent:
Tube | Standard Volume | Reagent 1 Volume (μL) | Concentration (mg/mL) |
Std.1 | 1 mL of Standard | - | 0.5 |
Std.2 | 500µL of Std.1 | 500 | 0.25 |
Std.3 | 500µL of Std.2 | 500 | 0.125 |
Std.4 | 500µL of Std.3 | 500 | 0.0625 |
Std.5 | 500µL of Std.4 | 500 | 0.03125 |
Std.6 | 500µL of Std.5 | 500 | 0.0156 |
Std.7 | 500µL of Std.6 | 500 | 0.0078 |
Blank | 0 | 500 | 0(Standard blank tube) |
Note: In the following procedure, 400 µL of standard is required for each standard tube (do not measure the absorbance of the standard directly at this step).
2.4. Pipette the reagents into 10 mL EP tubes as described in the table below.
Reagent (μL) | Test Tube | Blank Tube | Standard Tube |
Sample | 0.5 g | - | - |
Reagent 2 | 5000 | 5000 | - |
Incubate at 37°C in the dark. At 10 min and 3 h 10 min of the reaction, transfer 400 μL of the reaction mixture from each test tube and blank tube to perform the second-step reaction described below. For the second-step reaction, first add Reagent 3 and Reagent 4 to new 1.5 mL EP tubes according to the table below, then transfer 400 μL of the first-step reaction mixture into the EP tubes. Note: Reagent 3 and Reagent 4 must be added first, followed by the reaction mixture. The order must not be reversed! | - | ||
Reagent 3 | 400 | 400 | 400 |
Reagent 4 | 400 | 400 | 400 |
First-step reaction mixture | 400 | 400 | - |
Standard | - | - | 400 |
Mix thoroughly and incubate at room temperature for 20 min for color development. Transfer 1 mL of the mixture into a glass cuvette and measure the absorbance at 510 nm. The measurement should be completed within 20 min after color development. Record the absorbance at 10 min as A1 and the absorbance at 3 h 10 min as A2. Calculate ΔAtest = A1test - A2test, ΔAblank = A1blank - A2blank, ΔA = ΔAtest - ΔAblank , and ΔAstandard = Astandard - Astandard blank. (The standard curve and blank tube need only be run once or twice.)
3. Calculation of Root Activity
3.1. Preparation of the standard curve: Plot the standard curve based on the concentration of the standard tubes (x, mg/mL) and the absorbance ΔAstandard (y, ΔAstandard). Using the standard curve, substitute ΔA (y, ΔA) into the formula to calculate the sample concentration (x, mg/mL).
3.2. Calculation based on sample mass: Root activity is expressed as the amount of oxidized α-naphthylamine. Root activity [μg/(g·h)] = x × VReagent 2 × 10³ ÷ (W × T) = 1666.67 × x ÷ W 10³: unit conversion factor, 1 mg/mL = 10³ μg/mL; VReagent 2: volume of Reagent 2, 5 mL; W: root mass, g; T: reaction time, 3 h.
Notes
1. The order of adding Reagent 3, Reagent 4, and the sample into the EP tube must not be changed.
2. If the ΔA value is outside the range of the standard curve, either decrease the sample mass (or shorten the first-step reaction time) or increase the sample mass (or lengthen the first-step reaction time).
3. The measurement must be completed within 20 min after color development.
规格
规格或纯度
BioReagent, 生化试剂, 比色法
英文名称
Plant Root Activity Assay Kit (Naphthylamine, Colorimetric Method)
应用
细胞代谢
样品类型
植物
检测范围
0.0078-0.5 mg/mL
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
510nm
敏感性
Light-sensitive
检测时长
30 min
样本体积
400 μL
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
各组分在相应的储存条件下保质期1年。
组分与储存
P1515880 | 组分 | 物理外观 | 60 T | 储存条件 | 单次使用量 |
P1515880A | 试剂一 | 液体 | 20 mL | 2-8℃. | 按需 |
P1515880B | 试剂二 | 液体 | 250 mL | 2-8℃. | 按需 |
P1515880C | 试剂三 | 液体 | 25 mL | 2-8℃.避光保存. | 400 μL |
P1515880D | 试剂四 | 液体 | 25 mL | 2-8℃.避光保存. | 400 μL |
P1515880E | 标准品 | 液体 | 5 mL | 2-8℃.避光保存. | 按需 |
技术文档
高级数据
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技术文档和文章
溶液计算器
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