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. |
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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.
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