Hi, Z-Vad is used as inhibitor of apoptosis (general caspase inhibitor). I read that it has to be pre-incubated for an hour, before testing the drug of interest. But, can you please say whether z-vad has to be removed (after 1 hr exposure) before our drug is added to the cells, or can it be let in the media till the end of the experiment? I wanted to know, because my drug needs 72 hr exposure, and in the mean while the cells keep multiplying, so what will happen to the new cells? will they under go apoptosis or they are also get blocked?
yes, it can be there till the end of the experiment, both drug and fmk can be added together also.
if the fmk is present in sufficient amounts even the new cells will take up the inhibitor
You may also use an ubiquitous inhibitor which do not contain .fmk since fmk has its proper toxicity.
You may also test the fact of adding some more inhibitor each day if using a low concetration and compare to your experiment when you add the inhibitor early (for one our ) before the drug. and compare the results