I did proliferation assay twice and got similar IC50 : one after three days treatment with low dose drug and one with one day treatment with higher dose. Which IC50 shall I choose for my further assays as flow? The one with high dose short treatment or low dose long treatment ?
As a practical matter, I would choose the shorter assay to save time, as long as it gives the same result as the longer one. The length of time that is needed should depend upon the growth rate of the cells. You also need to consider the signal window in the assay. For a shorter growth period with the same starting number of cells, there will be a smaller window due to there being fewer cells at the end. If you are getting a reasonable signal-to-noise after one day, then go with that.
it depends on the speed of cell growth. Principally you must stop the trial before the 100% confluent cell of control.
good luck
I guess you can do 24,48 and 72 hours, enabling you to choose the time for flow assay (unless if you can run X-celligence, easier to observe at what point of time will be suitable). If in 24 hours you might not be able to differentiate death by necrosis or apoptosis. I would prefer at least 48 hours and make sure that the control does not reach 100% confluency. Probably around 80% at the end of the incubation. Just my opinion.
I would prefer 48hours, as it enables better to distinguish the diferences. However, some anticance drugs induce cell cycle arrest and then eventually cell death. In this later case longer observation period is necessary.
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