Hello, I´m having trouble with a luciferase reporter assay after estradiol stimuli, Some times I can measure the effect of estrogens and in other experiments i´m no able to measure the estradiol folding.

I´m doing something wrong, I had change the E2 stock and I did different preparations, but sometimes it works and others did NOT.

Can anyone help me? How do you prepare the betaE2 before the cell stimuli?

Thanks!!!