AP1 luciferase reporter...

Daniel O'Toole daniel.otoole at ucd.ie
Fri Nov 26 07:44:16 EST 2004


Sorry about the delay in replying. I've been using Genejuice
transfection reagent with HEK293 cells, and getting good TK-Renillin
control values, so I'm pretty sure my transfections are OK. I usually
transfect my similar NF-kB experiments for 24 hours and then treat
positive controls with IL1 or LPS for 6 hours, so I'm basically just
doing the same with a different reporter. I would expect that to be
long enough for accumulation of Luciferase protein. With one of the
AP1 reporters there appears to be plenty of signal, but it is
uninducible. Do you think a much shorter timeframe might help?
As an aside, would anybody have any experience with stably transfected
AP-1 reporter cell lines, which I've read of?
Thanks again.

> Daniel,
> 
> how is the setup of the experiment? Do you allow enough time for 
> accumulation of the reporter? What is the transfection efficiency of your 
> cells? Did you check the kinetics of reporter expression?
> 
> Regards,
> 
> Cornelius



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