45 Cells Lines suitable for Blasticidin S selection

February 7, 2013 Categories AG Scientific Blog

45 Cells Lines suitable for Blasticidin S selection

blasticidin_s_gene_selection_antibiotic_for_mammaliam_cell_linesBlasticidin S is an antibiotic used by scientists in bio-research to select cells modified by genetic engineering. It inhibits the growth of a wide range of prokaryotic and eukaryotic by interfering with protein synthesis cells.

Blasticidin S allows the selection and maintenance of cells expressing the blasticidin-resistance gene. Blasticidin S is a peptidyl nucleoside antibiotic isolated from the culture broth of Streptomyces griseochromogenes. It specifically inhibits protein synthesis in both prokaryotes and eukaryotes through inhibition of peptide-bond formation in the ribosomal machinery. It is used to select transfected cells carrying bsr or BSD resistance genes. Blasticidin S specifically inhibits protein synthesis in both prokaryotes and eukaryotes through inhibition of peptide-bond formation in the ribosomal machinery.

QUESTION: How do you determine Blasticidin sensitivity?

ANSWER:

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

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

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

d)   Determine the appropriate concentration of Blasticidin that kills the cells within 10-14 days after addition of the antibiotic.

QUESTION: What mammalian cell lines work with Blasticidin S?

ANSWER: See chart below.

Cell Type Selection Concentration ( µg/ml)
 10T1/2 cells  ~10 µg/ml
 293 HEK  3-10 µg/ml
 293T Lα  ~10 μg/ml
 A2780  ~5 µg/ml
 A549  2.5 to 10 μg/mL
 ASK  2.5-10 µg/ml
 B16  3-10 µg/ml
 CHO  5-10 µg/ml
 COS-1  3-10 µg/ml
 COLO 320DM  ~10 µg/ml
 CSII-EF-RfA  ~ 5 µg/ml
 Cell Type  Selection Concentration ( µg/ml)
 CV1-P  2.5-6 µg/ml
 D. discoideum coupled w/ Klebsiella aerogenes  ~ 10 µg/ml
 DH1  ~10 µg/ml
 DF-1  10 μg/ml
 EBV  ~5 µg/ml
 Giardia cells  ~75 μg/ml
 HAV  1-3 µg/ml
 HEK293T  2.5 to 10 μg/mL
 HeLa-S3  2.5 to 10 μg/mL
 HeLa  2.5 to 10 μg/mL
 HIV  10 µg/ml
 HMEC-1  2.5-6 µg/ml
 HMX44A  ~10 µg/ml
 HT1080  2.5 to 10 μg/mL
 FIV  10 µg/ml
 iPS  ~ 10 μg/ml
 JH10  ~10 µg/ml
 IMCD  ~5 μg/ml
 L929  2.5 to 10 μg/mL
 PC1.0  10-30 µg/ml
 MHOM/IL/80/Friedlin  10 µg/ml
 N2a  2.5 to 10 μg/mL
 NIH-3T3  2.5 to 10 μg/mL
 Nannochloropsis sp (algae)  ~5 μg/mL
 Plasmodium falciparum  2.5-5.0  μg/ml
 SHK-1  2.5-10 µg/ml
 S2 Drosophila  ~ 5 µg/ml
 S. cerevisiae  ~25 µg/ml
 TRE5-A  ~5 µg/ml
 T3-L1  2.5-10 µg/ml
 thraustochytrids  100 μg/ml
 TO  2.5-10 µg/ml
 U2OS  2.5 to 10 μg/mL

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