多酚氧化酶 (Polyphenol Oxidase, PPO)是自然界中分布极广的一种金属蛋白酶,普遍存在于植物、真菌、昆虫的质体中,甚至在土壤中腐烂的植物残渣上都可以检测到多酚氧化酶的活性,多酚氧化酶是一种蛋白体,在茶树生命活动和茶叶加工过程中参与一系列由酶促活动而引起的化学变化,故又被称为生物催化剂,该酶属于细胞木质素合成途径中间的关键酶,研究该酶可以探讨多种生物细胞发育过程中木质素沉积的代谢机理,为减少水果石细胞含量提高其品质提供依据。
多酚氧化酶 (PPO)提取试剂主要用于裂解植物组织,提取样品中的多酚氧化酶。该试剂仅用于科研领域,不适用于临床诊断或其他用途。
自备材料:
1、蒸馏水
2、离心管或试管、匀浆器或研钵、低温离心机
操作步骤 (仅供参考):
1、取新鲜的植物组织清洗干净,切碎。
2、按植物组织:多酚氧化酶 (PPO)提取试剂=3g: 6ml的比例,加入多酚氧化酶 (PPO)提取试剂,冰浴情况下充分捣碎或研磨。
3、10000g,4℃离心10-15min,留取上清液。
4、4℃冻存,用于多酚氧化酶 (PPO)的检测或其他用途。
注意事项:
1、待测样品中不能含有磷酸酶抑制剂,同时需避免反复冻融。
2、所测样本的值高于标准曲线的上限,应用提取试剂稀释样品后重新测定。
3、试剂开封后请尽快使用,以防影响后续实验效果。
4、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
Polyphenol Oxidase (PPO) is a metalloproteinase widely distributed in nature. It is generally present in the plastids of plants, fungi and insects, and PPO activity can even be detected in decaying plant residues in the soil. As a proteosome, PPO participates in a series of enzyme-catalyzed chemical reactions during the life activities of tea plants and the processing of tea leaves, hence it is also known as a biocatalyst. As a key enzyme involved in the intracellular lignin biosynthesis pathway, research on PPO can facilitate the exploration of metabolic mechanisms underlying lignin deposition during the developmental processes of various biological cells, providing a theoretical basis for reducing the content of stone cells in fruits and improving their quality.
The Polyphenol Oxidase (PPO) Extraction Reagent is primarily designed for lysing plant tissues and extracting PPO from samples. This reagent is intended for research use only and is not suitable for clinical diagnosis or any other applications.
Materials to Be Prepared by the User
1. Distilled water
2. Centrifuge tubes or test tubes, homogenizer or mortar and pestle, refrigerated centrifuge
Operating Procedures (For Reference Only)
1. Take fresh plant tissue, rinse it thoroughly, and cut it into small pieces.
2. Add the Polyphenol Oxidase (PPO) Extraction Reagent at a ratio of 3 g plant tissue: 6 ml reagent, then fully homogenize or grind the mixture under ice bath conditions.
3. Centrifuge at 10,000×g and 4°C for 10-15 minutes, and collect the supernatant.
4. Store the supernatant at 4°C for subsequent Polyphenol Oxidase (PPO) activity assays or other experimental applications.
Precautions
1. The sample to be tested must not contain phosphatase inhibitors, and repeated freeze-thaw cycles should be avoided.
2. If the measured value of the sample exceeds the upper limit of the standard curve, dilute the sample with the extraction reagent and perform the assay again.
3. Please use the reagent as soon as possible after opening to prevent it from affecting the results of subsequent experiments.
4. For your safety and health, wear a lab coat and disposable gloves during the operation.