I am concurrently trying to analyze the rate of protein synthesis under different siRNAs treatment followed by treatment with Puromycin (1 uM final concentration and 30 min exposure) after optimal incubation for knockdown of GOI (Gene of Interest). My control is Cyclohexamide treated cells which suppose to show a weak signal in respect to puromycin alone. As a readout, I employed western blotting, FACS, and ICC via using anti-Puromycin Antibody, clone 12D10 (Millipore_MABE343). I hope, the concentration of Puromycin is not lethal (cause total SHUT DOWN of protein in global level).

It would be great if someone can share some real-time experience of lab conduction and if possible share protocol specifically for cardiomyocytes.

Thank you very much. 

Here is the link of one of the most cited publication about this method:

http://www.nature.com/nmeth/journal/v6/n4/full/nmeth.1314.html