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AAT Bioquest

What should I consider when using protease inhibitors in the lab?

Posted March 21, 2024


Answer

There are several factors one should consider when using protease inhibitors in the lab. One thing to consider is to use a protease inhibitor cocktail. This cocktail protects proteins from various proteases, saving time and money compared to trying different inhibitors individually. Additionally, these cocktails are valued for their consistent and dependable performance. Inhibitor cocktails are typically formulated in stock solutions with a neutral pH, usually around 7. Since different inhibitors have distinct mechanisms, it's essential to identify the specific proteases to block before selecting the right inhibitor. This targeted strategy ensures effective protease inhibition. Another factor to consider is storage. Certain protease inhibitors can degrade rapidly when stored at room temperature or in the refrigerator for extended periods, particularly at their working concentrations. However, if stored correctly (with the manufacturer’s instructions) these inhibitors should maintain their efficacy until needed. It is also important to add a protease inhibitor just before lysing the cells. This prevents degradation. PMSF may require multiple additions during lysis to ensure continuous protease inhibition. Another consideration is whether to use a tablet, liquid, or powder form inhibitor. Tablets are popular because they offer convenience. Lastly, one should consider proper safety measures such as working in a fume hood, as certain inhibitors come as concentrated solutions in acetic acid or organic solvents.

Additional resources

Protease Inhibitors 101: How They Work and How to Use Them

Proteases

Amplite® Universal Fluorimetric Protease Activity Assay Kit *Green Fluorescence*