Hello everyone! I have to perform gene reporter assays in HEK293T to reveal if a gene I'm studying is regulated by NFkB. I am cloning the predicted NFkB binding regions into a vector driving luciferase expression. I have two questions:

1. Is it necessary to clone enhancer elements (i.e. immunoglobulin kappa light chain motifs) upstream the target gene promoter?

2. Is it fine to use the most common reported cytokine TNFalpha to activate NFkB pathway?

Thanks so much