Gibco™ Blasticidin S HCl (10 mg/mL)

Catalog No.	Quantity
A1113903	10 x 1 mL
A1113902	20 mL

Catalog No. A1113903 Supplier Gibco™ Supplier No. A1113903

Description

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

Blasticidin S is a nucleoside antibiotic isolated from Streptomyces griseochromogenes. It is a potent inhibitor of protein synthesis in both prokaryotic and eukaryotic cells. Blasticidin S acts quickly by inhibiting peptide bond formation by the ribosome.

• Used for prevention of contamination of cell cultures
• Recommended working concentration ranges 1–20μg/mL depending on cell line
• Provided as solution at 10mg/mL in 20 mM HEPES (pH 7.2–7.5)
• Supplied in 10 vials each containing 1mL.
• Resistance is conferred by expression of either one of two Blasticidin S deaminase genes: BSD or bsr

Cell Culture, Cloning, Drosophila S2 Cell Culture, Insect Cell Culture, Mammalian Cell Culture, Selection, Sf9 and Sf21 Cell Culture, Transfection, Transformation

Order Info

Shipping Condition: Dry Ice

Specifications

• Content And Storage: Storage conditions: -5°C to -20°C (protect from light)
  Shipping conditions: Dry ice
  Shelf life: 9 months from date of manufacture
• Cell Type: Eukaryotic Cells, Prokaryotic Cells
• Concentration: 10 mg/mL
• Culture Type: Mammalian Cell Culture, Insect Cell Culture
• Form: Liquid
• Product Type: Antibiotic
• Sterility: Sterile-filtered
• With Additives: HEPES
• Product Line: Gibco
• Quantity: 10 x 1 mL

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.

How can I decontaminate my cultures?

When an irreplaceable culture becomes contaminated, researchers may attempt to eliminate or control the contamination.

1. Determine if the contamination is bacteria, fungus, mycoplasma, or yeast. Read more here to view characteristics of each contaminant.
2. Isolate the contaminated culture from other cell lines.
3. Clean incubators and laminar flow hoods with a laboratory disinfectant, and check HEPA filters.
4. Antibiotics and antimycotics at high concentrations can be toxic to some cell lines. Therefore, perform a dose-response test to determine the level at which an antibiotic or antimycotic becomes toxic. This is particularly important when using an antimycotic such as Gibco Fungizone reagent or an antibiotic such as tylosin.

The following is a suggested procedure for determining toxicity levels and decontaminating cultures:

1. Dissociate, count, and dilute the cells in antibiotic-free media. Dilute the cells to the concentration used for regular cell passage.
2. Dispense the cell suspension into a multiwell culture plate or several small flasks. Add the antibiotic of choice to each well in a range of concentrations. For example, we suggest the following concentrations for Gibco Fungizone reagent: 0.25, 0.50, 1.0, 2.0, 4.0, and 8.0 µg/mL.
3. Observe the cells daily for signs of toxicity such as sloughing, appearance of vacuoles, decrease in confluency, and rounding.
4. When the toxic antibiotic level has been determined, culture the cells for two to three passages using the antibiotic at a concentration one- to two-fold lower than the toxic concentration.
5. Culture the cells for one passage in antibiotic-free media.
6. Repeat step 4.
7. Culture the cells in antibiotic-free medium for four to six passages to determine if the contamination has been eliminated.

What antibiotics do you offer to help control or eliminate cell culture contamination?

Please view the following page to browse the cell culture antibiotics we offer (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/antibiotics.html).

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

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