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Gibco™ Blasticidin S HCl, powder

Catalog No. R21001
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R21001 50 mg
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Catalog No. R21001 Supplier Gibco™ Supplier No. R21001
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Description

Bacterial selective antibiotic in concentration range of 50G–100μg/mL

Blasticidin S is a peptidyl nucleoside antibiotic isolated from Streptomyces griseochromogenes. It is a potent inhibitor of protein synthesis in bacteria and eukaryotes, while also active in fungi, nematodes, and tumor cells. Blasticidin S acts by blocking hydrolysis of peptidyl-tRNA induced by release factors and inhibits peptide bond formation. It is used as a selection agent in both mammalian and bacterial cells. The recommended working concentration ranges from 1 to 30 μg/mL depending on the cell line and 25–100 μg/mL for bacterial selection. Cell death occurs rapidly, and blasticidin-resistant stable mammalian cell lines can be generated in less than one week.

  • Blasticidin S acts quickly by inhibiting peptide bond formation by the ribosome
  • Used for prevention of contamination of cell cultures
  • Recommended working concentration ranges from 2 to 2μg/mL depending on the cell line
  • Resistance is conferred by expression of either one of two Blasticidin S deaminase genes: BSD (2) or bsr

Resistance to blasticidin S is conferred by BSR and BSD, isolated from Bacillus cereus K55-S and Aspergillus terreus respectively. The BSR resistance gene encodes blasticidin S deaminase, which catalyzes the conversion of blasticidin S to deaminohydroxyblasticidin S. Deaminohydroxyblasticidin S is a biologically inactive derivative of blasticidin S and does not interact with or inhibit prokaryotic or eukaryotic ribosomes. The BSD resistance gene also encodes a blasticidin S deaminase, which catalyzes a similar reaction to the BSR deaminase.

For bacterial selection purposes, the salt content of the LB medium must remain low (<90 mM) and the pH should not exceed 7.0 to maintain the activity of basticidin S. A kill curve is recommended in order to determine the minimum effective blasticidin S concentration to kill non-resistant cells.

Applications: Cell Culture, Cloning, Drosophila S2 Cell Culture, High Five™ Cell Culture, Insect Cell Culture, Mammalian Cell Culture, Selection, Sf9 & Sf21 Cell Culture, Transfection, Transformation

Order Info

Shipping conditions: Ambient

Specifications

Color White, off-white
Content And Storage Storage conditions: -5 to -30°C
Shipping conditions: room temperature
Shelf life: Lot Dependent
Cell Type Eukaryotic Cells, Prokaryotic Cells
Concentration 50 to 100 μg/mL
Culture Type Mammalian Cell Culture, Insect Cell Culture
Form Powder
Product Type Antibiotic
Sterility Sterile
With Additives HEPES
For Use With (Application) Bacterial Selection
Product Line Gibco
Quantity 50 mg
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Frequently Asked Questions (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.

For Research Use Only. Not for use in diagnostic procedures.