It turns out that my cells become 100 percent (from 70% confluent) confluent the after 4 days? Any suggestion On optimizing drug screening.?
Hi,
What is your concentration curve? How many concentrations?
I would try less confluent cells. Perhaps you could try 30% confluency and try your concentraion curve.
Thanks
At first I would suggest to grow only cells without your drug and find the right number for experiments. The cells should not overgrow during your experiments as you must have nice negative control at the end. Then to use serial dilutions of your tested compound, at least 6 concentrations. If you don't know anything about compound activity, you could make 10-fold dilutions from the highest concentration you have in cell growth medium. If your compound stock is in DMSO, you must to dilute it at least 200 times (DMSO concentration should not be higher than 0.5%). I prefer DMSO concentration not higher that 0.25%. In such way you will find approx. EC50 (effective concentration of compound that kills or inhibits growth of your cells by 50%). Then you may prepare more exact dilutions, like 2-fold dilutions. Previously calculated EC50 should be somewhere in the middle.
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