I am trying to establish a Gli reporter gene assay in Shh light 2 cells. For that I am using a dual luciferase reporter assay system. However the luminescence values obtained are only in the hundreds. I tried some modifications as well, but nothing helped.
Your suggestions will be of great help to me.
Can you please describe the experiment a bit as I am doing a a dual luciferase but getting numbers in the 100 thousands.
Hi Mohamed, I checked the firefly luciferase activity after treatment with Gli inhibitor and activator molecules. However in both the cases I get the luminescence values only in hundreds (~700). The Shh light 2 cells are stably transfected with GLI-responsive Firefly luciferase reporter and a constituitive Renilla-luciferase expression vector. I even tried transfecting other cell line with these plasmid but in that case also the luminescence remains the same. I followed many literature available and made the changes in my protocol (cell number, low serum media, incubation time, etc) but nothing helped.
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