Puromycin, Sterile 

Puromycin is an aminonucleoside antibiotic produced by Streptomyces alboniger. It inhibits protein synthesis by disrupting peptide transfer on ribosomes, causing premature chain termination during translation. It can kill most gram-positive bacteria and various animal or insect cells.

Resistance to Puromycin is conferred by the puromycin N-acetyl-transferase gene (pac) from Streptomyces alboniger. Therefore, Puromycin is a popular and effective antibiotic for selecting vectors bearing the pac gene in mammalian cell lines and E. coli.

-20°C; 2 years.

1. The working concentration of Puromycin varies with cell type, media, growth conditions and cell metabolic rate.

2. Before stable transfected cell lines can be selected, the optimal Puromycin concentration needs to be determined by performing a kill curve titration.

1) Seed the parental cell line in a suitable culture plate at a cell density of 20-25% and incubate the cells for 24 hours at 37°C.

2) Remove medium and then add medium with various concentrations of Puromycin and incubate at 37°C.

3) Refresh the selective medium every 2-3 days and observe the percentage of surviving cells over time.

4) Determine the lowest concentration of antibiotic that kills a large majority of the cells within 7-10 days. This concentration should be used for selection of a stable transfected cell line.