MG132 cell treatment and toxicity problem.

[cpatwardhan]

Hello Guys,
I use MG132 (a potent proteasomal inhibitor) to prevent/reduce the degradation of proteins by ubiquitin-proteasome machinery.
i am using 10uM of MG132 for 10 hrs but I observe 2 unwanted effects: 1) MG132 shows some cell toxicity and I lose ~30-40% of my cells. 2) MG132 itself targets my protein of interest and I do not see any effect of MG132 on the stability of my protein of interest.
I appreciate your suggestion.
Thanks,
Chaitanya

[youbati]

It may be too late for you, but for anyone being in trouble with MG132, you can try to treat your cells at a molarity ranging from 10µM to 20µM for ~4 hours. In my team, the routine treatment is 20µM, 4 hours, and we sometimes lower the concentration, depending on the cell line (20µM may still be toxic to some cells even with decreased treatment duration).

Otherwise, regarding the effect of MG132 itself towards a studied protein, one may want to use another inhibitor which is more proteasome activity-specific, such as lactacystin or epoxomicin.The drawback is that these inhibitors are far more expensive than MG132 is.