Does puromycin kill cells?

Puromycin quickly kills eukaryotic cells that do not contain the pac gene. Dying cells detach from the plates, allowing easy and early identification of transformant clones.

How long does G418 take to kill cells?

3. Replace the G418-containing medium every 2 – 3 days and examine cells for visual toxicity. Most non-transfected (non-resistant) cells will die within 10 days, leaving the transfected cells to expand.

Does neomycin kill mammalian cells?

Several reports suggest that neomycin is toxic for mammalian cell culture.

What is u2os cell?

Cells are positive for insulin-like growth factor I (IGF-I) and insulin-like growth factor II (IGF II) receptors and express a number of antigens, including blood type A, Rh+, HLA A2, Aw30, B12, Bw35, and B40(+/-). …

What should the concentration of puromycin be in a cell?

In mammalian cells, the recommended working concentration range for puromycin is 0.5 – 10 µg/ml. Different cell types and cell culture conditions may require different concentrations of selection antibiotic. Perform a kill curve to determine the optimal working concentra- tion for your experiment.

How long does it take for puromycin to kill a cell?

Puromycin has a fast mode of action, causing rapid cell death at low antibiotic concentrations. Adherent mammalian cells are sensitive to concentrations of 2 to 5 µg/ml, while cells in suspension are sensitive to concentrations as low as 0.5 to 2 µg/ml. Puromycin-resistant stable mammalian cell lines can be generated in less than one week.

How does puromycin work as an aminonucleoside?

Puromycin Puromycin is an aminonucleoside antibiotic produced by the bacterium Streptomyces alboniger. It inhibits protein synthesis by disrupting peptide transfer on ribosomes causing premature chain termination during translation. It is a potent translational inhibitor in both prokaryotic and eukaryotic cells.

How is Puromycin resistance conferred by the pac gene?

Resistance to puromycin is conferred by the puromycin N-acetyl-transferase gene ( pac) from Streptomyces. Puromycin has a fast mode of action, causing rapid cell death at low antibiotic concentrations.