reporter assay interpretation - repressor overexpression (Oct/12/2007 )
I need some help with the interpretation of my reporter assays. I wanted to see the effects of a repressor on my activation, but instead of repressing, the Luciferase values tripled!. B-gal control stayed the same. This is totally unexpected, as this repressor (SMRT) is not known to activate transcription.
What kind of weird effects must I be seeing???
I appreciate your help!
How much time you are incubating for B-gal assay???????//
What are the values for ur B-gal ?????????
U need to check these before normalising the luciferase values.
HI, thanks for replying!
I am following standarized protocols from my lab. Suspension cells, 5 ug B-gal, incubation period: 1 h after lysis. Values are very similar throughout the samples.