How to measure if forskolin has an affect? - (Nov/16/2005 )

Silly question ... what is a simple way to determine if my forskolin treatment of Cos7 cells results in activation of adenylate cyclase?

I am treating for both short (~4h) and long (24h) time periods with 0.1-10 µM forskolin.

The reason I am treating the cells with forskolin is to see if another protein is affected (which it isn't) but I would like to show that the forskolin acted appropriately so I can definitively say my protein is not a target.

Thanks,

AussieUSA.

Cell Signaling technology cells a phospho-specific antibody to phospho-PKA. On the data sheet they did flow cytometry showing increased FACS staining with the antibody when cells are treated with forskolin.

You could try using this antibody by flow to show your forskolin is working. This should be pretty simple to do but may not be the cheapest control experiment you could do.

Good Luck

Mountainman