Puromycin
| 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. |
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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) cells | 0.5 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) |
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