Cycloheximide-increase in protein levels???
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sciencemakerinthelab
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Cycloheximide-increase in protein levels???
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?
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?
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.
Neither of those seem likely, but as you say, this is a weird circumstance.
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