Hello, My lab is having great difficulty getting a PPRE luciferase reporter assay to work. I am able to get activation with ligand, but when we do anything else to manipulate the system, we are not getting any induction. This is very strange b/c typically we see coactivation with coactivators, etc. This system has completely stopped working for us. We have gotten new cells, new plasmids, new plasmid preps, new water, new media, you name, it, we've tried it, I think. It has been such a mystery as to why these luciferase assays are no longer showing induction as they were in the past. Any suggestions or ideas would be greatly appreciated.

Thanks

You can certainly sequence it to check whether the plasmid is fine. You can request again to the person who you got the plasmid from. I used this plasmid and got it from Dr. Peter Tontonoz (UCLA).

The problem isn't just one reporter. We are having trouble with all of our reporters and even the gal4 systems. It's the strangest thing.