Recently, I met a tricky system.
The proteins made from TNT.T7 reticulocyte lysate are sensitive to freezing and thawing steps.
I was assuming the denatured protein might be more stable than regular stuff.
But, even though I tested different ways to store them, the 2nd run for SDS-PAGE is completely different to 1st run.
I have a protease inhibitor inside. I tried to snap freeze my samples. I re-boiled my samples before loading.
Has anyone experienced something similar?