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乙醇酸氧化酶 (Glycolate Oxidase, GO)是乙醇酸循环的一种酶,在乙醇酸代谢循环中乙醇酸通过乙醇酸氧化酶的作用而变成乙醛酸。光合作用与呼吸作用是植物代谢的两大核心内容,前者是物质合成与能量储存过程,属于同化作用,为包括人来在内的几乎所有生物的生存提供物质来源和能量来源;后者是物质分解与能量释放过程,属于异化作用,为生命提供能量。通过测定样品中乙醇酸氧化酶活性,了解植物的光合和呼吸代谢的基本方法。
乙醇酸氧化酶(GO)提取试剂主要用于提取植物组织或果实中乙醇酸氧化酶。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:
研钵或匀浆器、纱布或滤纸、离心管或试管、离心机、pH计
操作步骤 (仅供参考):
1、取新鲜植物叶片,清洗干净,吸水纸吸干,称取18 g,加入18 ml预冷的GO提取试剂,冰浴情况下充分匀浆或研磨,经纱布或滤纸过滤,将滤液置于离心管或试管。
2、1000 g离心15 min,取上清液置于新的离心管或试管,调节pH值至5.4,4000 g离心15 min,取上清液。
3、按上清液: 蛋白沉淀剂=10 ml: 1.15 g的比例加入蛋白沉淀剂,不断混匀30 min,4000 g离心20 min,取上清液。
4、按上清液: 蛋白沉淀剂=10 ml: 0.6 g的比例加入蛋白沉淀剂,不断混匀30 min,4000 g离心20 min,弃上清液,留取沉淀即为乙醇酸氧化酶粗制品。
注意事项:
1、实验材料应尽量新鲜,如取材后不立即使用,应存于-20~-80 ℃。
2、试剂开封后请尽快使用,以防影响后续实验效果。
3、为了您的安全和健康,请穿实验服并戴一次性手套操作。
Glycolate Oxidase (GO) is an enzyme involved in the glycolate cycle. In the glycolate metabolic pathway, glycolate is converted to glyoxylate under the catalysis of glycolate oxidase. Photosynthesis and respiration are the two core processes of plant metabolism. Photosynthesis is the process of substance synthesis and energy storage, belonging to anabolism; it provides materials and energy for the survival of almost all organisms, including humans. Respiration is the process of substance decomposition and energy release, belonging to catabolism; it supplies energy for life activities. Determination of glycolate oxidase activity in samples is a basic method to analyze photosynthetic and respiratory metabolism in plants.
Glycolate Oxidase (GO) Extraction Reagent is mainly used for the extraction of glycolate oxidase from plant tissues or fruits.This reagent is for research use only and is not suitable for clinical diagnosis or other applications.
Materials Provided by User
Mortar or homogenizer, gauze or filter paper, centrifuge tubes or test tubes, centrifuge, pH meter
Protocol (For Reference Only)
1. Take fresh plant leaves, wash clean, blot dry with absorbent paper, and weigh 18 g. Add 18 mL of pre‑cooled GO Extraction Reagent, then homogenize or grind thoroughly in an ice bath. Filter through gauze or filter paper, and transfer the filtrate to a centrifuge tube or test tube.
2. Centrifuge at 1000×g for 15 min. Transfer the supernatant to a new centrifuge tube or test tube, adjust the pH to 5.4, then centrifuge at 4000×g for 15 min. Collect the supernatant.
3. Add Protein Precipitant at a ratio of supernatant: Protein Precipitant=10 mL: 1.15 g, mix continuously for 30 min, then centrifuge at 4000×g for 20 min. Collect the supernatant.
4. Add Protein Precipitant at a ratio of supernatant: Protein Precipitant=10 mL: 0.6 g, mix continuously for 30 min, then centrifuge at 4000×g for 20 min. Discard the supernatant; the pellet is the crude glycolate oxidase product.
Precautions
1. Experimental materials should be as fresh as possible. If not used immediately after collection, store at −20 to −80 ℃.
2. Please use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
3. For your safety and health, please wear a lab coat and disposable gloves during operation.
| G1509484 | Component | 2×500 mL | Storage |
| G1509484A | GO提取试剂 | 2×500 mL | 2-8℃. Store in the dark. |
| G1509484B | 蛋白沉淀剂 | 100 g | RT. Store in the dark. |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | G1509484 | |
| 分析证书 | G1509484 |
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