Selection Antibiotic Solutions

For stable cell line generation

Stable cell line generation allows for sustained expression of an exogenous gene which is commonly enriched and maintained through the use of selection antibiotics such as G418, Hygromycin B, Puromycin, etc. Resistance to these antibiotics is conferred through the expression of a positive selection gene: neomycin resistance (neo), hygromycin resistance (hph) and puromycin-N-acetyl-transferase (pac) genes, respectively.

Selection markers can be delivered using the same plasmid (in cis) that contains the gene of interest, or on a separate plasmid (in trans) that needs to be co-transfected with the plasmid containing the gene of interest. For a detailed protocol, please visit our Stable cell line generation web page.

Mirus offers high quality cell culture grade selection antibiotics with the following features:

Ease of use - Sterile filtered ready-to-use stock solutions for stable cell line generation

High Purity - Greater than 90% purity determined by HPLC

 High Potency - In combination with high purity ensures better selection at optimal concentrations

Figure 1. G418 Sulfate for Stable Cell Line Generation. G418 Sulfate is routinely used for positive selection of cells that express the neo gene encoding neomycin phosphotransferase. This occurs as only cells that have successfully integrated the neo gene can express the positive selection marker. The optimum concentration for selection is cell-type dependent; performing a kill curve is recommended with each new cell type or different antibiotic lot. Recommended concentration for the selection of mammalian cells that are resistant to G418 is 100-2000 µg/ml for mammalian cells. 

Figure 2. Hygromycin B for Stable Cell Line Selection.Hygromycin B is an aminoglycoside antibiotic that is used for the positive selection and maintenance of mammalian cells transfected with the E. coli  hygromycin resistance gene.  This occurs as only cells that have successfully integrated the hygromycin resistance gene can express the positive selection marker. The optimum concentration for selection is cell-type dependent; performing a kill curve is recommended with each new cell type or different antibiotic lot. Recommended concentration for the selection of mammalian cells that are resistant to Hygromycin B is 100-500 µg/ml for mammalian cells.

Figure 3. Puromycin for Stable Cell Line Selection. Puromycin is an aminoglycoside antibiotic that is used for the positive selection and maintenance of mammalian cells transfected with vectors that express puromycin-N-acetyl-transferase (PAC). This occurs as only cells that have successfully integrated the PAC gene can express the positive selection marker. The optimum concentration for selection is cell-type dependent; performing a kill curve is recommended with each new cell type or different antibiotic lot. Recommended concentration for the selection of mammalian cells that are resistant to Puromycin 0.5 - 10 µg/ml for mammalian cells.