Questions Related to V8 Protease
1) What is the pH and buffer system used?
V8 protease is typically dissolved in a buffer with a pH range of 8.0 to 8.5 at 37°C. The common buffers used are 50 mM NH4HCO3 or Tris-HCl. The concentration can be adjusted between 1 to 10 mg/mL depending on the application.
2) Storage Conditions
Store the enzyme at -20°C. It is recommended to avoid repeated freeze-thaw cycles. After up to five cycles, the enzyme should still retain more than 80% of its original activity.
3) Application
V8 protease is widely used in protein analysis, including structural studies and sequence determination. It is particularly effective for cleaving proteins at the C-terminal side of glutamic acid residues.
4) Specific Activity
The specific activity of V8 protease is approximately 20 U/mg when measured using Z-Phe-Leu-Glu-4-nitroanilide as a substrate at 25°C. At 37°C, the activity increases significantly when casein is used as a substrate, reaching around 500 U/mg.
Additional Notes:
This enzyme is known for its high specificity and stability under controlled conditions. It is commonly used in biochemical research, especially in the field of proteomics and protein digestion protocols. Always ensure that the working solution is freshly prepared and stored properly to maintain optimal performance.
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