Generally my drug has IC-50 value around 10ug/ml. which concentration should I use for time dependent western experiments? 10ug/ml or 15ug/ml or 20ug/ml or 40 ug/ml? which is best and why?
Trisha,
Your question on mechanisms of apoptosis is too generalistic and therefore a bit difficult to understand and address. The word 'mechanism' has too broad meaning and I have no idea what you have in your mind. What are you studying? Capases? mitochondriall function? cell signaling? transcription factors? link between apoptosis and autophagy.
Do you see, how helpless we become to understand your objectives.
In general, people use two to three concentrations to test. At least two, one low concentration at which 10% cells die and the other at 10 ug/ml. If you increase your concentration, your cells will die and you won't have cells/proteins available for your assays.
For example, if you are studying MAPKinases, you need cells that are still alive or in the process of dying. Hope this helps a bit. Next time, please be specific while framing these questions. Some of the answers to your questions also found reading other published work as how they have approched and how many doses they have used etc.
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