I'm going to use cycloheximide to inhibit the synthesis of a membrane protein in HEK-293T cells, but I'm not sure the right time that needed to treat cells without influencing its viability. I used the concentration of 100uM for 21h, which was mentioned in a similar article, cells would die a lot. I'm not sure whether the medium should be replaced after treating the cells for some time or should it be remained in the medium until the harvest of cells. Please tell me if you have experience on using this drugs. ☺Thanks a lot~
Cycloheximide the most common laboratory reagent used to inhibit protein synthesis. It blocks the elongation phase of eukaryotic translation. It binds the ribosome and inhibits eEF2-mediated translocation . Cycloheximide is supplied as a lyophilized powder and used at 5-50 µg/ml conc.
Thank you sir Madhukar (@ Madhukar Baburao Deshmukh ) , I have tested the toxicity of cycloheximide by using different concentrations and followed up the viobility changes at different time points yesterday. The results showed that a concentration of 100uM （≈90ug/ml）did not cause cell death at 18h, which is somehow different from my previous experiments. I'll do more tests to figure it out. Thank you again.
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