Hygromycin B is an aminoglycosidic antibiotic that inhibits protein synthesis by disrupting translocation and promoting mistranslation at the 80S ribosome. Because it uses a different mode of action than Gibco Geneticin, Blasticidin S, or Gibco Zeocin, it’s perfect for dual-selection experiments when used in conjunction with another selection agent.

Resistance to Hygromycin B is conferred by the E. coli hygromycin resistance gene (hyg or hph). The concentration for selection ranges from 100-1000 µg/ml (typically 200 µg/ml) and should be optimized for each cell line.

Formula: C20H37N3O13
FW: 527.5 g/mole

Table 1 - Selection agents and resistant vectors

Cell types
Conc. (μg/ml)*
Invitrogen vectors carrying resistance marker
pcDNA5 vectors (Constitutive mammalian expression)
pIND/Hygro vector (Ecdysone-Inducible mammalian expression)
Jurkat T-Cell
pSecTag2/Hygro vector (Secreted mammalian expression)
S2 Drosophila
pREP vectors (Episomal mammalian expression)
  pCoHygro (Selection vector for DES)

* The optimal concentration for selection of your cell line should be determined using kill curves.