After getting no bands on my Westerns, I now have too many! I have bands both bigger and smaller than the predicted size of my protein. I was looking through old posts and saw this is not an uncommon problem - and one old post suggested adding urea to the loading buffer to reduce dimerisation.
So, how much urea exactly do I add? And will any urea do? And why does it work?
Also, I saw several replies suggesting longer blocking - but would this not reduce all antibody binding? I know it's called non-specific binding but the antibody must be recognizing something in order to bind to the protein.