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
| N1518233 | Component | 500 mL | Storage | | N1518233A | Nitrate Reductase Extraction Reagent
| 500 mL | 2-8℃. Store in the dark. | | N1518233B | PMSF | 1 mL | -20℃. Store in the dark. |
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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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