I am planning to determine the IC50 dose of tamoxifen for different cell lines.
I plate the cells on day 1 and add tamoxifen in a range of different concentrations after 24 hours. From that time point on, how long should I incubate the cells before measuring viability? 48 hours? 72? Or even longer? Thanks in advance for your advices!
In my experience you have to take into account of the doubling time of the cell line in order to verify the effects also on cell proliferation. 24 hours could not be sufficient. In my opinion you have to test at least 3 exposure time 24, 48 and 72 hours. You could "jump" 48 hours if this time is not so relevant and introduce an Acute exposure of 6 hours. Moreover you have to indicate the time of your IC50 (for example 24h IC50, 72h IC50).
For cells in suspension you could perform an easy test with counting chamber and trypan blue at different time collecting a small amount of volume from the dishes.
If you are interesting only on cell viability you could use a time-course assays using some specific probes (such as one from Promega) that you will add to the culture medium and read the "fluorescence/luminescence" at the desired time point.
In my experience you have to take into account of the doubling time of the cell line in order to verify the effects also on cell proliferation. 24 hours could not be sufficient. In my opinion you have to test at least 3 exposure time 24, 48 and 72 hours. You could "jump" 48 hours if this time is not so relevant and introduce an Acute exposure of 6 hours. Moreover you have to indicate the time of your IC50 (for example 24h IC50, 72h IC50).
For cells in suspension you could perform an easy test with counting chamber and trypan blue at different time collecting a small amount of volume from the dishes.
If you are interesting only on cell viability you could use a time-course assays using some specific probes (such as one from Promega) that you will add to the culture medium and read the "fluorescence/luminescence" at the desired time point.
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