Generally speaking, protease inhibitor refers to the combination of a protein enzyme and some groups, which will decrease enzyme activity without changing the nature of the enzyme protein.
Among the application of protease inhibitor, there are many questions and doubts during experiment. So this post will give you some solutions to these questions.
Question 1:
Is there a need to add cocktail protease inhibitor in protein extraction?
Example:
Someone is doing the experiment on extracting proteins from organization for WB. And his lysates contains sodium pyrophosphate, β-glycerophosphate, EDTA, Na3VO4, leupeptin, and other inhibitors, which can inhibit the degradation of proteins effectively. So if is there a need for him to add cocktail protease inhibitor?
Solution:
As for this question, others express that adding cocktail protease inhibitors is according to proteins you use in your experiment. Some proteins, which do not need inhibitors, can be extracted. Some proteins only need PMSF (Phenylmethanesulfonyl fluoride). Proteins, which can be degraded easily, would better be added to some cocktail protease inhibitor.
Question 2:
When we are doing an experiment on dialysis, what kind of protease inhibitors should we add in order to prevent protein degradation?
Solution:
There are a lot of protease inhibitors among this field. Some inhibitors can just inhibit one kind of protein enzyme activity, so some labs will turn to protease inhibitors for Serine sites. If we want to get a broad spectrum of inhibitory effect, the cocktail protease inhibitors can offer us help. This kind of inhibitor is formed by all kinds of inhibitors. So it can offer the best effect in inhibiting. Of course, some proteins do not need protease inhibitors, if this experimentation is processed under low temperature conditions. And then the protein precipitation may lie in front of us.
These two questions are very common in using the protease inhibitors. Here we introduce the solutions to them. And we hope this can help you a lot.
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