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硝酸还原酶 (Nitrate Reductase, NR)是一种氧化还原酶,可分为参与硝酸盐同化的同化型还原酶和催化以硝酸盐为活体氧化的最终电子受休的硝酸盐呼吸异化型(呼吸型)还原酶。硝酸还原酶是植物氮素代谢中氮素同化的关键酶,该酶与作物吸收利用氮肥有关,对作物的产量和质量有影响,因此可以把硝酸还原酶的活力当作营养诊断养、农田施肥或作物育种的生理生化指标。
硝酸还原酶 (NR)提取试剂主要用于裂解植物组织,提取样品中的硝酸还原酶。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:
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自备材料:
1、蒸馏水
2、离心管或试管
3、匀浆器或研钵
4、低温离心机
操作步骤 (仅供参考):
1、取0.5 g植物组织清洗干净,切碎,置于-20 ℃冰箱30 min。
2、配制硝酸还原酶提取工作液:取出硝酸还原酶提取试剂和PMSF,恢复至室温,按硝酸还原酶提取试剂: PMSF=499: 1的比例混合,即为硝酸还原酶提取工作液 (NR Lysis Buffer),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: NR Lysis Buffer=0.5 g: 4 ml的比例,加入预冷的硝酸还原酶提取工作液,冰浴情况下充分匀浆或研磨。
4、4000 g,4 ℃离心15-20 min,留取上清液即为硝酸还原酶粗提液,4 ℃保存,用于硝酸还原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、硝酸还原酶容易失活,提取和测定时均应操作迅速,尽量在4 ℃操作。
3、如果测定植物样本,取样最好在晴天进行,最好提前一天施些硝态氮肥,取样部位应一致。
4、待测样品中不能含有磷酸酶抑制剂,同时需避免反复冻融。
5、所测样本的值高于标准曲线的上限,应用丙酮酸脱羧酶提取工作液稀释样品后重新测定。
6、试剂开封后请尽快使用,以防影响后续实验效果。
7、为了您的安全和健康,请穿实验服并戴一次性手套操作。
Nitrate Reductase (NR) is an oxidoreductase. It can be classified into assimilatory reductase involved in nitrate assimilation and dissimilatory (respiratory) nitrate reductase that catalyzes nitrate respiration with nitrate as the final electron acceptor for vital oxidation. Nitrate Reductase is a key enzyme for nitrogen assimilation in plant nitrogen metabolism. This enzyme is related to the absorption and utilization of nitrogen fertilizer by crops and affects crop yield and quality. Therefore, the activity of Nitrate Reductase can be used as a physiological and biochemical index for nutritional diagnosis, farmland fertilization, or crop breeding.
Nitrate Reductase (NR) Extraction Reagent is mainly used for lysing plant tissues and extracting Nitrate Reductase from samples.This reagent is for research use only and is not suitable for clinical diagnosis or other applications.
Components
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Materials Provided by User
1. Distilled water
2. Centrifuge tubes or test tubes
3. Homogenizer or mortar
4. Refrigerated centrifuge
Protocol (For Reference Only)
1. Take 0.5 g of plant tissue, clean thoroughly, mince, and place in a −20 °C refrigerator for 30 min.
2. Prepare Nitrate Reductase Extraction Working Solution:
Take Nitrate Reductase Extraction Reagent and PMSF to room temperature. Mix at a ratio of Nitrate Reductase Extraction Reagent: PMSF=499:1 to obtain Nitrate Reductase Extraction Working Solution (NR Lysis Buffer). Use immediately after preparation; prolonged storage reduces the inhibitory efficiency of PMSF.
3. Mix minced plant tissue with pre‑cooled NR Lysis Buffer at a ratio of plant tissue: NR Lysis Buffer=0.5 g: 4 mL. Homogenize or grind thoroughly in an ice bath.
4. Centrifuge at 4000 × g, 4 °C for 15-20 min. The supernatant is the crude Nitrate Reductase extract. Store at 4 °C for NR detection or other analyses.
Calculation
Crude Enzyme Extract Yield (mL/g)=Supernatant Volume (mL)/Sample Weight (g) × 100%
Precautions
1. Use fresh experimental materials whenever possible. If not used immediately, store at −20 to −80 °C.
2. Nitrate Reductase is prone to inactivation. Perform extraction and assay rapidly, preferably at 4 °C.
3. For plant samples, sampling is recommended on sunny days, with nitrate fertilizer applied one day in advance, and sampling position should be consistent.
4. Test samples must not contain phosphatase inhibitors; avoid repeated freeze‑thaw cycles.
5. If sample readings exceed the upper limit of the standard curve, dilute the sample with Pyruvate Decarboxylase Extraction Working Solution and re‑assay.
6. Use the reagent promptly after opening to avoid compromising experimental performance.
7. For your safety and health, wear a lab coat and disposable gloves during operation.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | N1518233 |
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