I work with E. coli and I want to examine the stability of my protein of interest via Western blot. For this purpose I am planning to grow my cultures under conditions that induce the synthesis of my protein, harvest 1 ml sample and then add tetracycline for a certain period of time and harvest another sample. The idead is that tetracycline stops protein synthesis and if my protein is stable ithe amont of protein in both samples should be the same. If the protein is unstable its amount should decrease after tetracycline treatment.
My question is has anyone tried this before and if yes what tetracycline concentration would you recommend me? I suppose it should be higher than the usual tetracycline concentration used for selection (12 ug/mL) but so far I was unable to find an exact concentration for such a treatment in literature. Thank you very much!