Questions Related to V8 Protease
1. What is the pH and buffer system used?
The V8 protease is typically dissolved in a buffer solution with a pH range of 8.0 to 8.5 at 37°C. Common buffers include 50 mM NH4HCO3 or Tris-HCl. The concentration can be adjusted between 1-10 mg/ml depending on the application.
2. Storage Conditions
It should be stored at -20°C. Repeated freeze-thaw cycles (up to 5 times) are acceptable, but it's recommended to minimize such cycles to maintain optimal activity. The enzyme should retain over 80% of its original activity under these conditions.
3. Application
V8 protease is commonly used for protein structure and sequence analysis. It cleaves proteins at the carboxyl side of glutamic acid residues, making it useful in various biochemical studies and peptide mapping techniques.
4. Specific Activity
The specific activity of V8 protease is approximately 20 U/mg at 25°C when using Z-Phe-Leu-Glu-4-nitranilide as a substrate. At 37°C, the activity increases significantly when casein is used as the substrate, reaching around 500 U/mg. This makes it a powerful tool for enzymatic digestion in research settings.
Additional Notes: Always ensure proper handling and storage to preserve enzyme integrity. For best results, use fresh solutions and avoid prolonged exposure to high temperatures or repeated freezing.
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