I am establishing a Gli reporter assay in Shh light 2 cells using a Dual luciferase reporter assay system, but I am getting really low luminescence values. However, the same protocol works perfectly fine for other genes in different cells.
What modifications should I consider in the case of Shh light 2 cells?
Hi, did you try the same gene in other cells)? I wonder that is the problem in cell or gene(or construct).
is your transfection control (assuming you are using beta galactosidase) also low? if so, your transfection is the culprit.
Hi Mert and Matthew, The Shh Light 2 cells are stably transfected. However, I also tried transient transfection in another cells but still the luminescence is low and I used commercial construct and in house construct.
You transfected to other cells and it was still low? Maybe it is all about its basal level maybe there is problem in construct. Did you try to induce it with some activators to see any changing (upregulation)?
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