Hygromycin B

Hygromycin B is an antibiotic used to select prokaryotic and eukaryotic cells. It works by interfering with protein synthesis. Cloning of the resistance genes and fusion with eukaryotic promoters has resulted in vectors to select for resistance in both prokaryotic and eukaryotic cells.

Mode of action
Interferes with protein synthesis.

Mechanism of resistance
Resistance genes are kinases that inactivate Hygromycin B through phosphorylation.

Stability
sensitive to high concentrations of acid, but short-term exposure to dilute acids tolerated;
 * >1y at –20°C
 * >2y at +4°C (http://www.hygromycin.com/service.htm)
 * 3m at RT
 * >1m at 37°C

Usage notes
The working concentration for selection varies with cell type, media, growth conditions and cell metabolic rate. Recommended concentration for resistant cells varies between 25 and 1000 mg/ml. You need to determine your optimum concentration experimentally. For some typical values by cell types see the table below.

For more information on working concentrations: