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Cycloheximide-increase in protein levels???

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Cycloheximide-increase in protein levels???

Postby sciencemakerinthelab » Sun Jan 27, 2013 2:56 pm

Hi everyone,

In order to find out the half life of my protein of interest, I treated my cells with cycloheximide(CHX), which is an eukaryotic translation inhibitor, through different time intervals. Then, I looked at the protein amounts by Western Blot. However, I saw that as I left them in medium containing CHX more, the levels of my protein of interest increased. How could it be possible?
sciencemakerinthelab
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Postby Darby » Sun Jan 27, 2013 7:03 pm

It would suggest that some production of your protein is not translation-driven. Can another protein be enzymatically converted into the one you're looking at? Could there be a storage-release issue, with the stored version not showing up until it's coverted to the active form?

Neither of those seem likely, but as you say, this is a weird circumstance.
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