Blasticidin S HCl, powder - FAQs

哺乳动物细胞的稳转筛选过程中可一起使用哪些种类的抗生素（Geneticin，Zeocin，潮霉素B，杀稻瘟素和嘌呤霉素）？

我们所有的抗生素（Geneticin，Zeocin，潮霉素B，杀稻瘟素和嘌呤霉素）均可共用于多重稳转细胞系的建立。不过，需要针对每一种抗生素组合来获取杀菌曲线，因为当与其他抗生素联用时，细胞对某一特定抗生素的灵敏性会出现增加的趋势。

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.