Pyruvate decarboxylase (PDC), also known as α‑carboxylase, is a carboxylase acting on α‑keto acids. Pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) are key enzymes in the ethanol fermentation pathway. Both are up‑regulated under hypoxic conditions. PDC activity is generally lower than ADH activity, making it the rate‑limiting enzyme for ethanol synthesis. The accumulation of metabolites from the anaerobic respiration pathway is toxic to cells, affecting mitochondrial structure and the activities of enzymes involved in the tricarboxylic acid cycle.
Pyruvate Decarboxylase (PDC) Extraction Reagent is mainly used for lysing plant tissues and extracting pyruvate decarboxylase from samples.This reagent is for research use only and is not suitable for clinical diagnosis or other applications.
Components
| P1518228 | Component | 500 mL | Storage | | P1518228A | Pyruvate Decarboxylase Extraction Reagent
| 500 mL | 2-8℃. Store in the dark. | | P1518228B | 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. Collect plant tissue, clean thoroughly, and mince.
2. Prepare working solution of Pyruvate Decarboxylase Extraction Reagent:
Take Pyruvate Decarboxylase Extraction Reagent and PMSF to room temperature. Mix at a ratio of Pyruvate Decarboxylase Extraction Reagent: PMSF=499: 1. Use immediately after preparation; prolonged storage reduces the inhibitory efficiency of PMSF.
3. Mix minced plant tissue with pre‑cooled working solution at a ratio of plant tissue: working solution=1 g: 4 mL. Homogenize or grind thoroughly in an ice bath.
4. Centrifuge at 12000 × g, 4 °C for 20 min. The supernatant is the crude pyruvate decarboxylase extract. Store at 4 °C for PDC 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. Test samples must not contain phosphatase inhibitors; avoid repeated freeze‑thaw cycles.
3. If sample readings exceed the upper limit of the standard curve, dilute the sample with PDC extraction working solution and re‑assay.
4. Use the reagent promptly after opening to avoid compromising experimental performance.
5. For your safety and health, wear a lab coat and disposable gloves during operation.
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