Time course experiment on protein level - (Apr/01/2021 )

What is the purpose from doing a time course experiment on protein level?

For example adding a drug in one particular concentration at different time points.

-sun shine 411-

What do you think the purpose is? Why might time of drug addition matter in an infectious process?

-bob1-

I think to see when the effect of the drug is going to appear, and to show the maximum effect and when the effect will disappear (minimum effects). That will help in the future with an experiment to show the perfect effect because we know the maximum effect will appear at specific time.

-sun shine 411-

Exactly.

-bob1-

One more question, please. If I prepared the drug for a time course, let say my time course experiment is 24 hours, and I prepared a stock from the drug and at each time point doses the cells from this stock. So keep the stock in the fridge 4C and pre-warmed every time I dose the cells. Is that OK? The goal of this is to prevent the difference between wells. Or do I need to prepare a fresh drug every time I am dosing the cells?

Thanks

-sun shine 411-

How stable is the drug? If it has no/little degradation after storage at 24 hours, then you should be fine to keep it under the conditions you mention. If it degrades in that time, then you will have to prepare fresh each time.

If you do experience degradation, you may be able to aliquot and freeze the drug - though in this case it would be best to aliquot beforehand, freeze all aliquots, then thaw one for the first time point and one at each subsequent time point, so that they all undergo the same freeze/thaw cycle.

-bob1-