苹果酸脱氢酶 (Malate Dehydrogenase, MDH)是合成苹果酸的关键酶之一，催化苹果酸和草酰乙酸 (OAA)的相互转化，参与众多生理代谢途径如TCA循环C4循环脂肪酸的氧化呼吸作用氮同化等，因此MDH在植物的生长发育中发挥着重要作用，广泛存在于线粒体、细菌细胞膜上，为三羧酸循环中的一种酶，由于酶的来源不同，其某些性质也不尽相同，MDH在细胞多种生理活动中扮演着重要的角色，包括线粒体的能量代谢、苹果酸-天冬氨酸穿梭系统、活性氧代谢和抗病性等。
苹果酸脱氢酶 (MDH)提取试剂主要用于裂解植物组织，提取样品中的苹果酸脱氢酶。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
产品组成：
| A1518214 | Component | 500 mL | Storage | | A1518214A | 苹果酸脱氢酶提取试剂 | 500 mL | 2-8℃. Store in the dark. | | A1518214B | PMSF | 1 mL | -20℃. Store in the dark. |
|
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、取植物组织清洗干净，切碎。
2、配制苹果酸脱氢酶提取工作液：取出苹果酸脱氢酶提取试剂和PMSF，恢复至室温，按苹果酸脱氢酶提取试剂: PMSF=499: 1的比例混合，混匀，即配即用，不易久置，否则蛋白酶抑制剂PMSF的效率会有所下降。
3、按植物组织: 苹果酸脱氢酶提取工作液=1 g: 4 ml的比例，加入预冷的苹果酸脱氢酶提取工作液，冰浴情况下充分匀浆或研磨。
4、12000 g，4 ℃离心20 min，留取上清液即为苹果酸脱氢酶粗提液，4 ℃保存，用于苹果酸脱氢酶的检测或其他用途。
计算：
样品粗酶液获得率 (ml/g)=上清液体积 (ml)/样品质量 (g)×100%
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、待测样品中不能含有磷酸酶抑制剂，同时需避免反复冻融。
3、所测样本的值高于标准曲线的上限，应用苹果酸脱氢酶提取工作液稀释样品后重新测定。
4、试剂开封后请尽快使用，以防影响后续实验效果。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Malate Dehydrogenase (MDH) is one of the key enzymes for malate synthesis, which catalyzes the reversible conversion between malate and oxaloacetate (OAA). It participates in numerous physiological metabolic pathways, including the TCA cycle, C4 cycle, fatty acid oxidation, respiration, nitrogen assimilation, etc. Therefore, MDH plays an important role in plant growth and development. MDH is widely distributed in mitochondria and bacterial cell membranes, and is an enzyme in the tricarboxylic acid cycle. Its properties may vary depending on the enzyme source. MDH is involved in a variety of cellular physiological processes, including mitochondrial energy metabolism, malate‑aspartate shuttle system, reactive oxygen species metabolism, disease resistance, etc.
Malate Dehydrogenase (MDH) Extraction Reagent is mainly used for lysing plant tissues and extracting malate dehydrogenase from samples.This reagent is for research use only and not suitable for clinical diagnosis or other applications.
Components
| A1518214 | Component | 500 mL | Storage | | A1518214A | MDH Extraction Reagent
| 500 mL | 2-8℃. Store in the dark. | | A1518214B | PMSF | 1 mL | -20℃. Store in the dark. |
|
Materials Provided by User
1. Distilled water
2. Centrifuge tubes or test tubes, homogenizer or mortar, refrigerated centrifuge
Protocol (For Reference Only)
1. Take plant tissues, wash clean, and cut into pieces.
2. Prepare MDH Extraction Working Solution:
Take out MDH Extraction Reagent and PMSF, equilibrate to room temperature. Mix at a ratio of MDH Extraction Reagent: PMSF = 499: 1. Prepare fresh before use; do not store for a long time, otherwise the inhibitory efficiency of PMSF will decrease.
3. Add pre‑cooled MDH Extraction Working Solution at a ratio of plant tissue: working solution=1 g: 4 mL, then homogenize or grind thoroughly in an ice bath.
4. Centrifuge at 12000×g, 4℃ for 20 min. The supernatant is the crude MDH extract. Store at 4℃ for MDH detection or other experiments.
Calculation
Crude Enzyme Extract Yield (mL/g) = Supernatant Volume (mL) / Sample Weight (g) × 100%
Precautions
1. Experimental materials should be as fresh as possible. If not used immediately, store at −20 to −80℃.
2. Test samples should not contain phosphatase inhibitors, and repeated freeze‑thaw cycles should be avoided.
3. If the measured value exceeds the upper limit of the standard curve, dilute the sample with MDH Extraction Working Solution and re‑assay.
4. Please use the reagent as soon as possible after opening to avoid affecting experimental results.
5. For your safety and health, please wear lab coat and disposable gloves during operation.
ZJ26F0434119
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