I face a problem the last weeks I wanted to discuss with u.I extract total protein from cultured cells and I determine the concentration of the lysates by Bradford.It looks that my samples are protein-rich. The problem is that after carrying out the western blot, ponceau as well actin show that the protein amount I supposed to load is much lower.
In few words Bradford shows much protein and western only few. However I use other control proteins and these show that western blot and transfer work well.
I checked the reagents of my lysis buffer and the concentrations are compatible with bradford. So I was wondering what it may go wrong and interferes with bradford and how I could solve the problem.Any idea?
Thanks in advance for any help.