Protease Inhibitors - (Apr/30/2006 )
Hello, I'm rather new in the area of proteomics.
I would like to query about protease inhibitors..
Are examples of these inhibitors Aprotinin and Leupeptin?
Why are protease inhibitors added into the lysis buffer?
What are its uses, advantages and disadvantages?
Thanks.
I would like to query about protease inhibitors..
Are examples of these inhibitors Aprotinin and Leupeptin?
Why are protease inhibitors added into the lysis buffer?
What are its uses, advantages and disadvantages?
Thanks.
Yes,
Protease inhibitors are added to the lysis buffer to protect the protein of the animal and plant tissues or cells during extraction.
It is used to protect the protein from serine, cysteine and metalloprotease which are found in the animal or plant tissues.
Sorry, I do not know the disadvantage.
UHm, ok thanks.. can anyone add more details?
I don't know of any disadvantages, either, except you have to spend money to get them...but it's well worth it. they prevent degradation. if you've ever spent days and weeks working through a protocol, only to get to the end and find out that all your hard work was wasted because your samples degraded and you have to start over, then you understand the value of protease inhibitors 
Minnie did a pretty good job...specifically, what would you like to know? you may choose to go to this link...click on a few and check out the details...for precise specific mechanisms you would probably want to do a pubmed search by name of inhibitor; there are quite a few and their actions are varied...but basically they inhibit proteases, which are pretty much everywhere and can wreak havoc on your extracts and lysates
the only disadvantage would be if you were working with a protease that is irreversibly inhibited by one of these compounds.
Is there anyway of redeeming an experiment if you forget to add protease inhibitors?
sometimes your protein of interest won't be affected by the proteases if you work quickly enough or if it is not susceptible to attack by the prevalent proteases.
Is there anyway of redeeming an experiment if you forget to add protease inhibitors?
sometimes your protein of interest won't be affected by the proteases if you work quickly enough or if it is not susceptible to attack by the prevalent proteases.
THanks a lot. There are a bunch of proteins I'll be looking at, but I hope I worked fast enough to minimize degradation. I also tried to keep everything on ice as often as possible. They're just whole cell lysates that I'm running out, so if I add sample buffer and boil them, that should inactivate any proteases?
most proteases will be denatured by boiling but i worked with one that was still fully active after 15 minutes of boiling (hot block set to 105C).
good luck.