Puromycin is an aminonucleoside antibiotic produced by the bacterium Streptomyces alboniger. It inhibits protein synthesis by disrupting peptide transfer on ribosomes causing premature chain termination during translation. It is a potent translational inhibitor in both prokaryotic and eukaryotic cells. Resistance to puromycin is conferred by the puromycin N-acetyl-transferase gene (pac) from Streptomyces. Puromycin has a fast mode of action, causing rapid cell death at low antibiotic concentrations. Adherent mammalian cells are sensitive to concentrations of 2 to 5 µg/ml, while cells in suspension are sensitive to concentrations as low as 0.5 to 2 µg/ml. Puromycin-resistant stable mammalian cell lines can be generated in less than one week.

Table 1 - Suggested Puromycin Concentration

Cell Line Concentration [µg/ml] Reference
HT1080 (human fibrosarcoma)0.2–0.4
1
2
J Biol Chem 282:29314–29322 (2007)
J Virol 82:3320–3328 (2008)
Cancer Res 68: 1417–1426 (2008)
HeLa (human cervical cancer)1-2
2
2
Genes Cells12:397–406 (2007)
Mol Cell Biol 28:4104–4115 (2008)
PNAS 105:16490–16495 (2008)
HEK293 (human embryonic kidney)0.25
2
3
Nucleic Acids Res 36:e83 (2008)
Mol Cell Biol 28:4104–4115 (2008)
Mol Cell Biol 27:4708–4719 (2007)
Hep G2 (human hepatocellular carcinoma)0.3
2
2
J Biol Chem 283:21462–21468 (2008)
Mol Cell Biol 28:4104–4115 (2008)
J Biol Chem 283:708–715 (2008)
Human embryonic stem (ES) cells0.5
1
5
Nucleic Acids Res 36:e148 (2008)
Stem Cells 26:850–863 (2008)
Stem Cells 26:2245–2256 (2008)
MCF-7 (human breast cancer)0.45
1
2
RNA 13:1375-1383 (2007)
Cancer Res 68:1319–1328 (2008)
Mol Cell Biol 28:4104–4115 (2008)
H1299 (non-small cell lung carcinoma)1
1.5
2.5
Nucleic Acids Res 35:e17 (2007)
J Biol Chem 283:33394–33405 (2008)
Proc Natl Acad Sci USA 105:1937–1942 (2008)
MDA-MB-231 (human breast cancer)0.5
1.5
5
Mol Cell Biol 28:997–1006 (2008)
Mol Cell Biol 27:324 – 339 (2007)
Mol Cell Biol 28:687–704 (2008)
A549 (lung cancer)1.5
1.5
Mol Cell Biol 27:324–339 (2007)
J Biol Chem 283:33394–33405 (2008)