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HELP:Pretreated with inhibitor before 1 h? - (Nov/20/2011 )

Hi. Signal pathway inhibitors, such as LY294002 for PI3K/Akt, are often used to explore the role of the signal axis. It's usually described in the paper that "pretreated with LY294002 for 1 h before other treatment", then what's the meaning of the "pretreated for 1 h"?
Should I change the medium after 1 h treatment with the inhibitor and before other treatment, or just add the other treatment without discarding the inhibitor?

Since it's my first topic in the forum, I'm not sure I choose the right field for publication.

-note1988-

You got to make sure the pretreated drug action should remain until the end of the time point of the later treated drug..So in this case some drug action would revert in mins to hours once you take it out, so better you pipette out the pretreated medium in a falcon, add the second drug, mix and again add into the flask.. i recommend this rather than wasting another aliquot of drug for the same treatment.

-GNANA-

GNANA on Mon Nov 21 08:53:57 2011 said:


You got to make sure the pretreated drug action should remain until the end of the time point of the later treated drug..So in this case some drug action would revert in mins to hours once you take it out, so better you pipette out the pretreated medium in a falcon, add the second drug, mix and again add into the flask.. i recommend this rather than wasting another aliquot of drug for the same treatment.


Thanks for your help. That's quit helpful. I think maybe I should performed in these two ways, and then detected the targeting gene expression or activation. Therefore, to choose a better method.

-note1988-