Geneticin (G418) is an aminoglycoside antibiotic similar in structure to Gentamicin produced by Micromonospora rhodorangea. Geneticin is widely used for selection of eukaryotic cells, stably transfected with neomycin resistance genes (neo) as well as maintenance of the (neor ) phenotype of resistant cells. Geneticin is toxic to bacteria, yeast, protozoa, helminths and mammalian cells. It can also be used for elimination of contaminating fibroblasts from mixed cultures.
Geneticin (G418) blocks polypeptide synthesis by inhibiting protein elongation by binding irreversibility to 80S ribosomes both in prokaryotic and eukaryotic cells. This activity can be inactivated by the bacterial aminoglycoside phosphotranferases APH(3’)II and APH(3’)I encoded by genes on transposons Tn5 and Tn601 (903) respectively. The cells acquire resistance to Geneticin (neo) as a result of transfection of neomycin resistance genes (neo) from transposon Tn5 or Tn601 and enable the cells to grow in media containing Geneticin (G418).
This selection strategy can be used on almost any cell type. The effective concentration of geneticin varies according to the cell type, medium used, growth conditions, cell’s metabolic rate and position in the cell cycle. We recommend determining optimal concentrations of antibiotic required to kill your host cell line by treating the cells with several concentrations ranging from 100μg/ml to 1mg/ml. Cells can escape selection if the antibiotic is used at too low concentrations or if the plating density is too high. Ideally cell death of control cells occurs within one week after addition of antibiotic allowing colonies of resistant cells to form by 10-14 days.
相關產品列表
購物說明：