Blasticidin S HCl, powder - FAQs

Which of your antibiotics (Geneticin, Zeocin, Hygromycin B, Blasticidin, and Puromycin) can be used together for stable selection in mammalian cells?

All of our antibiotics (Geneticin, Zeocin, Hygromycin B, Blasticidin, and Puromycin) can be used together for making multiple stable cell lines. However, kill curves will need to be performed for each combination of antibiotics since sensitivity to a given antibiotic tends to increase when combined with other antibiotics.

What are the recommended concentrations of antibiotics to use for selection in prokaryotes and eukaryotes?

For best results, optimal concentrations for selection should be determined empirically in each unique experiment through dose response curves. However, to get a general idea of concentrations that have worked for individual cell types, please click on the following url: http://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/selection.html or type in “Selection Antibiotics” into our main search on www.thermofisher.com.

What is the mode of action on the following antibiotics: Blasticidin, Geneticin (G418), Hygromycin, and Zeocin?

Blasticidin: Nucleoside Inhibits protein synthesis in prokaryotic and eukaryotic cells by interfering with peptidyl transfer reaction of protein synthesis, causing early termination of translation.

Geneticin (G418): Aminoglycoside Blocks protein synthesis in mammalian cells by interfering with ribosomal function.

Hygromycin: Aminocyclitol Inhibits protein synthesis by disrupting translocation and promoting mistranslation.

Zeocin: Intercalates with DNA and cleaves it.

What is the optimal pH of low salt LB for LB + blasticidin plates?

We recommend a pH of 7 or less and half the normal amount of NaCl in your LB media or plates.

See the following paper for details on optimal conditions: Yamaguchi et al (1965) Inhibition of Protein Synthesis by Blasticidin S. Journal of Biochemistry (Tokyo) Volume 57: pp 667-677.

How long can Blasticidin be stored at 4 degrees C after thawing? Does the unused portion have to be discarded after thawing?

Blasticidin is stable for 6 months when stored at 4 degrees C. Discard remaining material after this time.

What solvent can be used to dissolve Blasticidin?

Blasticidin can be dissolved in water or acetic acid. Make stocks of 5-10 mg/ml.