PMA / Ionomycin, Revised & Simplified Question, Please Help! - (Aug/21/2008 )

Hi,

I am doing a dual luciferase system cotransfection in Jurkat cells. The PMA and Ionomycin seems to be negatively affecting my cells, based on a crashing of activity of both my experimental and control reporters. My unstimulated cells (no PMA/Ionomycin) give me better results by 2 orders of magnitude. PMA Iono should be increasing activity of my experimental plasmid tremendously.

Here is my question:

Can someone tell me *start to finish* how to dilute PMA and ionomycin, including suggested storage, what to dilute with, etc? Any hints, etc etc.

Thanks,

Mike