AP1 luciferase reporter...

Cornelius Fritsch c.fritsch at dkfz.de
Thu Nov 11 01:58:28 EST 2004


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


"Daniel O'Toole" <daniel.otoole at ucd.ie> schrieb im Newsbeitrag 
news:63f68427.0411100641.5085756 at posting.google.com...
> Hi all,
> I was wondering if anybody has any experience of using AP1-luciferase
> reporter plasmids. I've been trying to get Clontech and Stratagene
> ones to work.
> The Clontech one seems to give hardly any signal and only a few fold
> induction by MEKK. There's the opposite porblem with the Stratagene,
> which seems to be activated maximally by serum in the medium. I've
> tried using serum free (eg Optimem) but then I have trouble inducing
> the signal again.
> Basically, I'm assuming I can get IL-1 or LPS or something to activate
> AP1 (not just MEKK), but I can't seem to get either AP1 reporter to
> measure this. Any suggestions?
> Thanks 





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