过氧化物酶(peroxisome，POD)是以过氧化氢为电子受体催化底物氧化的酶，主要存在于细胞的过氧化物酶体中，以铁卟啉为辅基，可催化过氧化氢，氧化酚类和胺类化合物，具有消除过氧化氢和酚类、胺类毒性的双重作用，该酶属于细胞木质素合成途径中间的关键酶，可用于探究植物发育过程中木质素积累的代谢规律，为调控果实石细胞生成、改善果实品质提供检测依据。

植物过氧化物酶 (POD) 检测试剂盒 (愈创木酚微板法) 检测原理是以愈创木酚,又称 2-甲氧基酚作为底物，在酶促反应的最适条件下采用每隔一定时间测定产物生成量的方法，于酶标仪 470 nm 处测定吸光度，以吸光度变化所需酶量进行计算，主要用于植物组织的裂解液或匀浆液等样品中内源性的过氧化物酶活性，尤其适用于测定水果中过氧化物酶活性。该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。

组分货号	组分名称	物理外观	100T	储存	单次测试使用量
P1521768A	POD 裂解液	液体	100.0 mL	2-8 ℃.	145.0 μL
P1521768B	POD 检测液	液体	10.0 mL	2-8 ℃. 避光	按需
P1521768C	POD 氧化剂	液体	1.0 mL	2-8 ℃. 避光	按需

自备材料：

酶标仪、台式离心机、水浴锅/恒温培养箱、可调节式移液枪及枪头、研钵或匀浆器、96孔板 、2M 硫酸 (可选) 、30 -50目筛、冰和蒸馏水。

使用方法

1. 准备样品

1) 植物样品：取新鲜植物组织，按照组织质量（g）：提取液体积（mL）为1：10 的比例用研钵进行冰浴研磨，制备成10%植物匀浆液，如 0.1 g 植物叶片，加入 1 mL 提取液，用研钵进行冰浴研磨。12000 rpm，4℃离心 10 min，取上清，即为含有 POD 的样本，置冰上测定。

2) 高活性样品：如果样品中含有较高活性的过氧化物酶，可以使用POD 裂解液进行恰当的稀释。

注：如果提高研磨效率，可加入适量石英砂（试剂盒不提供，自备）辅助研磨；提取的样本如需贮存，建议按需分装后-80℃贮存，两周内使用，尽量避免反复冻融。

2. POD工作液配制

1) 取适量的 POD 氧化剂和 POD 检测液 (按照检测样品个数计算)，按 POD 氧化剂：POD检测液=1：14 混合，即为 POD工作液，即配即用，不宜久置。

3. 样本测定

1) 酶标仪开机预热 30 min 以上，调节波长 470 nm。

2) 检测前，试剂盒中的试剂平衡至室温。

3) 取 96 孔板，按照下表设置对照孔孔、测定孔。对照孔、测定孔中为同一待测样品，但对照孔中是提前加热煮沸 5 min 的样品；溶液应按照顺序依次加入，并注意避免产生气泡，如果样品中的 POD 活性过高，可以减少样品用量或适当稀释后再进行测定，样品的检测最好能设置 2 平行孔，求平均值。

加入物 (µL)	对照孔	测定孔
待测样品	5 (提前煮沸 5 min)	5
POD 裂解液	145	145
POD 工作液	100	100

4) POD 测定：立即以酶标仪，以对照孔为对照，测定 470 nm 处测定孔的吸光度(A_测定0)；37℃准确孵育 3 min 后，立即加入 5 μL 2M硫酸终止反应 (备选方案)，以对照孔为对照，以酶标仪测定 470 nm 处测定孔的吸光度( A_测定1) ，或使用酶标仪连续检测3 min动力循环 (37℃)进行实时数据测定 (推荐)。

注：加入2M 硫酸终止反应不是必须步骤，可 37℃准确孵育 3 min 后，以对照孔为对照，直接以酶标仪测定 470 nm 处测定孔的吸光度( A_测定1 )。

4. POD 活性计算 POD 活性单位的定义：在该实验条件下，每 1 min 吸光度变化 0.01 所需酶量为一个活性单位。

1) 组织样本 POD(U)={( A_测定1 - A_测定0)× V₁ }/(W× Vₛ ×0.01×t)

A_测定1=孵育 3 min 后测定孔的吸光度值

A_测定0 =加入POD工作液后测定孔的吸光度值

W=组织样本的重量(g) V₁=提取酶液的总体积(mL

Vₛ=测定时所用酶液体积(mL)

t=反应时间 (min）

2) 液体样本 POD(U)=( A_测定1 - A_测定0)/(0.01×t)

A_测定1 =孵育 3min 后测定孔的吸光度值

A_测定0 =加入 POD Assay buffer 工作液后立即测定的测定孔吸光度值

t= 反应时间 (min）

注意事项：

1. 待测样品中不能含有酶抑制剂，同时需避免反复冻融。

2. POD 酶提取时，注意低温操作，防止酶活性。

3. 以煮沸的酶液为对照时，酶要充分失活。

4. POD 氧化剂具有一定腐蚀性，请小心操作。

5. POD 检测液易挥发，请密闭保存，否则检测效率下降。

6. 本产品仅供科学研究使用。请勿用于临床、诊断、食品、化妆品检测等用途。

7. 为了您的安全和健康，请穿实验服并戴一次性手套操作。

Product Introduction

Peroxidase (POD) is an enzyme that catalyzes substrate oxidation using hydrogen peroxide as the electron acceptor. It is mainly located in cellular peroxisomes and contains iron porphyrin as its prosthetic group. It can catalyze hydrogen peroxide to oxidize phenolic and amine compounds, exerting dual functions of eliminating hydrogen peroxide and detoxifying phenols and amines. As a key intermediate enzyme in the cellular lignin biosynthetic pathway, investigations of this enzyme help elucidate the metabolic mechanism of lignin deposition during cell development in various organisms, and provide a basis for reducing cellular lignin content in fruits to improve fruit quality.

The detection principle of the Plant Peroxidase (POD) Assay Kit (Guaiacol Microplate Method) is as follows: guaiacol (also named 2-methoxyphenol) serves as the substrate. Under optimal enzymatic reaction conditions, the amount of reaction product is measured at fixed time intervals. The absorbance is read at 470 nm using a microplate reader, and enzyme activity is calculated based on the change in absorbance over time. This kit is used to determine endogenous peroxidase activity in samples such as lysates or homogenates of plant tissues, and is particularly suitable for assaying peroxidase activity in fruits. This kit is for research use only; it is not intended for clinical diagnosis or any other applications.

Component No.	Component Name	Appearance	100T	Storage	Quantity Per Test
P1521768A	POD Lysis Buffer	Liquid	100.0 mL	2-8 ℃.	145.0 μL
P1521768B	POD Assay Buffer	Liquid	10.0 mL	2-8 ℃.Store in the dark.	As needed
P1521768C	POD Oxidant	Liquid	1.0 mL	2-8 ℃.Store in the dark.	As needed

Materials to be Supplied by User

Microplate reader, bench-top centrifuge, water bath/constant temperature incubator, adjustable pipettes and pipette tips, mortar or homogenizer, 96-well plate, 2 M sulfuric acid (optional), 30-50 mesh sieve, ice and distilled water.

Operating Procedure

1. Sample Preparation

1) Plant samples: Weigh 0.5-1.0 g plant tissue or mesocarp of fruit, add to 5 mL pre-cooled POD lysis buffer, then grind or homogenize. Centrifuge at 10000 g for 15-20 min at 4 °C. Collect the supernatant as crude POD extract. Store at -20 °C for subsequent peroxidase assay.

2) High-activity samples: If samples contain high peroxidase activity, appropriately dilute the sample with POD lysis buffer.

Note: To improve grinding efficiency, an appropriate amount of quartz sand (not supplied in the kit, to be prepared by the user) may be added to assist grinding. If the extracted samples need to be stored, it is recommended to aliquot them as required and store at -80 °C, use within two weeks, and avoid repeated freeze-thaw cycles as much as possible.

2. Preparation of POD Working Solution

1) Take adequate volumes of POD oxidant and POD detection solution (calculated based on the number of test samples). Mix POD oxidant : POD detection solution = 1 : 14 to prepare POD working solution. Prepare immediately before use; do not store for extended periods.

3. Sample Measurement

1) Power on the microplate reader and preheat for ≥30 min, set wavelength to 470 nm.

2) Before assay, equilibrate all kit reagents to room temperature.

3) Prepare a 96-well plate and set up control wells and test wells according to the table below. Control wells and test wells use the same test sample. The sample in control wells is pre-boiled for 5 min. Add solutions sequentially and avoid bubble formation. If POD activity in the sample is excessively high, reduce sample loading volume or dilute the sample prior to measurement. It is recommended to set 2 replicate wells for each sample and calculate the average value.

Reagent (µL)	Control well	Test well
Test sample	5 (pre-boiled for 5 min)	5
POD lysis buffer	145	145
POD working solution	100	100

4) POD measurement: Immediately read the absorbance of test wells at 470 nm on the microplate reader using control wells as blank reference, record as A_test0. Incubate accurately at 37 °C for 3 min. Option 1 (stop reaction): Add 5 μL of 2 M sulfuric acid to terminate the reaction. Read absorbance of test wells at 470 nm with control wells as reference, record as A_test1. Option 2 (recommended): Perform continuous kinetic reading for 3 min at 37 °C on microplate reader for real-time data acquisition. Note: Adding 2 M sulfuric acid is optional. After 3 min incubation at 37 °C, absorbance A_test1 of test wells at 470 nm can be directly measured with control wells as reference without reaction termination.

4. Calculation of POD Activity Definition of POD activity unit: Under the assay conditions, one unit (U) is defined as the amount of enzyme required to cause an absorbance change of 0.01 per minute.

1) Tissue sample POD activity (U): ={( A_Test1- A_Test0)× V₁ }/(W× Vₛ ×0.01×t)

Note:

A_Test1= absorbance of test well after 3 min incubation.

A_Test0 = absorbance of test well immediately after adding POD working solution.

W= weight of tissue sample (g)

V₁ = total volume of enzyme extract (mL)

Vₛ= volume of enzyme solution used in assay (mL)

t= reaction time(min）

2) Liquid sample POD activity (U): =( A_Test1 - A_Test0)/(0.01×t)

Note:

A_Test1 = absorbance of test well after 3 min incubation

A_Test0 = absorbance of test well immediately after adding POD working solution

t=reaction time (min）

Precaution

1. Test samples must not contain enzyme inhibitors; repeated freeze-thaw cycles should be avoided.

2. Perform extraction of POD enzyme solution under low temperature to prevent loss of enzyme activity.

3. When using boiled enzyme solution as control, ensure complete enzyme inactivation.

4. POD oxidant is corrosive; handle with caution.

5. POD Assay Buffer is volatile. Keep tightly sealed during storage; otherwise detection performance will decrease.

6. This product is for research use only. Not for clinical diagnosis, food testing or cosmetic testing.

7.Wear lab coat and disposable gloves for your safety and health during operation.

规格

规格或纯度

BioReagent, 比色法, 适用于分析

英文名称

Plant Peroxidase (POD) Activity Assay Kit (Guaiacol, Micro Method)

应用

植物过氧化物酶, 细胞代谢

检测方法

分光光度法

定量&半定量

定量

检测仪器

酶标仪

检测波长

470nm

敏感性

Light-sensitive

检测时长

3 min

样本体积

5 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

组分货号	组分名称	物理外观	100T	储存	单次测试使用量
P1521768A	POD 裂解液	液体	100.0 mL	2-8 ℃.	145.0 μL
P1521768B	POD 检测液	液体	10.0 mL	2-8 ℃. 避光	按需
P1521768C	POD 氧化剂	液体	1.0 mL	2-8 ℃. 避光	按需

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:

技术文档和文章

植物逆境代谢网络怎么分析？GABA、脯氨酸、可溶性糖与抗氧化系统联动评价

目录：技术文章

探索主题:

ROS 渗透调节抗氧化系统 SOD CAT MDA 植物逆境 GABA检测脯氨酸检测可溶性糖检测 POD APX 植物胁迫响应

How to Analyze Plant Stress Metabolic Networks? Integrated Evaluation of GABA, Proline, Soluble Sugars, and the Antioxidant System

目录：技术文章

探索主题:

ROS Antioxidant system SOD CAT MDA POD APX GABA detection Plant Stress Proline Detection Soluble Sugar Detection Osmotic Regulation Plant Stress Response

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

浓度

体积

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

体积 1 (V1)

浓度 2 (C2)

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

质量（小瓶）

所需的复溶浓度

常见问题

什么是「BioReagent」？

「BioReagent」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。

本产品应如何储存？

请于2–8 °C、避光条件下保存。本品对光敏感，请保存在原始的避光或棕色容器中。

本产品如何运输？

本产品采用湿冰冷藏运输。收货后请立即拆包，并按上述储存条件保存。

如何获取批次专用COA？

每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。

需要帮助选择级别？

我们的级别选择指南涵盖了目录中所有的变体的纯度、稳定剂状态和应用适用性。

查看 BioReagent 级别指南 → 查看 Suitable for Analysis 级别指南 → 查看 Colorimetry 级别指南 →

大包装与定制包装 需要更大包装或定制规格？请联系我们的团队获取专属报价。