Blasticidin S HCl solutionA nucleoside antibiotic supplied at 10 mg/mL in HEPES Buffer for use as a selection antibiotic.

Blasticidin S HCl solution (CAS 3513-03-9)

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Anwendungen A nucleoside antibiotic supplied at 10 mg/mL in HEPES Buffer for use as a selection antibiotic.
CAS Nummer: 3513-03-9
Reinheit: ≥98%
Molekulargewicht: 458.90
Summenformel: C17H26N8O5·HCl
Supplemental Information: This is classified as a Dangerous Good for transport and may be subject to additional shipping charges.
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Blasticidin S HCl solution is supplied ready to use at 10 mg/mL in HEPES Buffer and Sterile Filtered. Blasticidin S is a nucleoside antibiotic isolated from Streptomyces griseochromogenes which inhibits protein synthesis in both prokaryotic and eukaryotic cells. Blasticidin S acts quickly by inhibiting peptide bond formation by the ribosome. Resistance is conferred by expression of one of two Blasticidin S deaminase genes: BSD from Aspergillus terreus or bsr from Bacillus cereus. These deaminases convert Blasticidin S to a nontoxic deaminohydroxy derivative.

Blasticidin S is used for the prevention of contamination of cell cultures. The recommended working concentration ranges from 1–20 µg/mL depending on the cell line.

Blasticidin S may also be used as a bacterial selection antibiotic in the concentration range of 50–100 µg/mL. To maintain the activity of Blasticidin S for selection purposes, the salt content of the LB medium must remain low (<90 mM) and the pH should not exceed 7.0.


Literaturhinweise

1. Yamaguchi, H., Yamamoto, C. and Tanaka, N. 1965. Inhibition of protein synthesis by blasticidin S. I. Studies with cell-free systems from bacterial and mammalian cells. J. Biochem. 57(5): 667-677. PMID: 4953866
2. Izumi, M., Miyazawa, H., Kamakura, T., Yamaguchi, I., Endo, T. and Hanaoka, F. 1991. Blasticidin S-resistance gene (bsr): a novel selectable marker for mammalian cells. Exp. Cell Res. 197(2): 229-233. PMID: 1720391
3. Kimura, M., Takatsuki, A. and Yamaguchi, I. 1994. Blasticidin S deaminase gene from Aspergillus terreus (BSD): a new drug resistance gene for transfection of mammalian cells. Biochim. Biophys. Acta. 1219(3): 653-659. PMID: 7948022
4. Kim, H., Ishidate, T., Ghanta, K.S., Seth, M., Conte, D., Shirayama, M. and Mello, C.C. 2014. A co-CRISPR strategy for efficient genome editing in Caenorhabditis elegans. Genetics. 197(4): 1069-1080. PMID: 24879462

Gebrauch :
Caution

Toxic if swallowed. Contains Blasticidin S. Avoid contact with skin and eyes. Wear suitable protective clothing and gloves. In case of accident or if you feel unwell, seek medical advice immediately.



Storing Blasticidin

• Do not subject Blasticidin solution to multiple freeze-thaw cycles (do not store in a frost-free freezer).

• When stored properly at –5°C to –20°C, Blasticidin is stable for up to 9 months. Store medium containing Blasticidin at 4°C for up to 2 weeks.

• Upon thawing, use what you need and discard any unused portion.



Blasticidin Selection in E. coli

For selection of Blasticidin-resistant E. coli, use Low Salt LB medium (10 g Tryptone, 5 g sodium chloride, 5 g yeast extract) containing 50–100 µg/mL Blasticidin. Optimize the Blasticidin concentration for the bacterial strain being used. If a lawn of bacteria forms on the Low Salt LB plate instead of individual bacterial colonies, increase the Blasticidin concentration in the plates to 100 µg/mL.

Note: The salt concentration of the medium must remain low (<90 mM). Failure to lower the salt content of your LB medium inhibits Blasticidin, and prevents selection unless a higher Blasticidin concentration is used.



Blasticidin Selection in Yeast

The concentration of Blasticidin required for selection in yeast varies depending on the species, strain, and type of medium used. Use a range of 25–300 µg/mL Blasticidin for selection in yeast. Standard protocols recommend performing a kill curve for each species, strain, and medium used to determine the appropriate Blasticidin concentration to use for selecting resistant cells.



Blasticidin Selection in Mammalian Cells

The Blasticidin concentration required for selection in mammalian cells varies depending on the cell line used. Use 2–10 µg/mL Blasticidin for selection in mammalian cells. Standard protocols recommend performing a kill curve as described below to determine the appropriate Blasticidin concentration to use for selecting resistant cells.



Determining blasticidin sensitivity

1. Plate cells at approximately 25% confluency. Prepare a set of 6 plates. Allow cells to adhere overnight.

2. The next day, substitute culture medium with medium containing varying concentrations of Blasticidin (e.g. 0, 2, 4, 6, 8, 10 µg/mL Blasticidin).

3. Replenish the selective media every 3–4 days, and observe the percentage of surviving cells.

4. Determine the appropriate concentration of Blasticidin that kills the cells within 10–14 days after addition of the antibiotic.
Formulierung :
10 mg/mL in HEPES Buffer, Sterile Filtered
Aggregatzustand :
Liquid
Erhaltene Form :
Streptomyces griseochromogenes
LAGERUNG: :
Store at -20° C
Ausschließlich für Forschungszwecke. Nicht Geeignet für Verwendung in Diagnostik oder Therapie.
Deutsche WGK :
2
Transport :
UN 3172, Class 6.1, Packing group III
PubChem CID :
25078183
MDL-Nummer :
MFCD02091640
SMILES :
CN(CC[C@H](CC(=O)N[C@H]1C=C[C@@H](O[C@@H]1C(=O)O)N2C=CC(=NC2=O)N)N)C(=N)N.Cl

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Blasticidin S HCl solution  Publikationen

Lesen Sie wie andere Anwender Blasticidin S HCl solution eingesetzt haben. Klicken Sie auf den jeweiligen Eintrag um den entsprechenden PubMed-Abstract abzurufen .

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PMID: # 30838870  Wu, M.|Feng, Y.|Ye, GX.|Han, YC.|Wang, SS.|Ni, HF.|Wang, FM.|Gao, M.|Lv, LL.|Liu, BC.| et al. 2019. Am. J. Physiol. Renal Physiol. 316: F1006-F1015.

PMID: # 30881116  Chiang, KC.|Huang, ST.|Wu, RC.|Huang, SC.|Yeh, TS.|Chen, MH.|Hsu, JT.|Chen, LW.|Kuo, SF.|Chueh, HY.|Juang, HH.|Hung, SI.|Yeh, CN.|Pang, JS.| et al. 2019. Cancer Manag Res. 11: 1893-1905.

PMID: # 30034634  Barone, G.|Arora, A.|Ganesh, A.|Abdel-Fatah, T.|Moseley, P.|Ali, R.|Chan, SY.|Savva, C.|Schiavone, K.|Carmell, N.|Myers, KN.|Rakha, EA.|Madhusudan, S.|Collis, SJ.| et al. 2018. Oncotarget. 9: 29508-29524.

PMID: # 30538136  Thwaites, DT.|Carter, C.|Lawless, D.|Savic, S.|Boyes, JM.| et al. 2018. Blood.

PMID: # 31576091  World J. Gastroenterol. 25: 5434-5450.

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