I am trying to block protein synthesis in living cells with using cycloheximide. I am monitoring this by labeling proteins with 35S-methionine during CHX treatment. I've tried several concentrations (0.5-100 ug/ml) and it seems to work to some extent (no difference between 0.5-20 ug though). Unfortunately, the forward transport of the protein I am investigating seems to be blocked as well (I am not getting an EndoH resistant band anymore, which is the case with no CHX).
Could the cycloheximide powder, that we stored at room temp for 2-3 years, have "gone bad" and therefore, has low efficiency and kills the cells as a side effect?
We have experienced similar results as you comment Zeynep. And, even though it is not explicit in the flask we suspect that somehow it gets bad after two or three years.
Dear Bart and Maria,
thank you for your responses. I haven't found out unfortunately why the compound did not work.
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