after a serial of alanine mutation on some region of interest of a protein, i noticed substitution of alanine of some amino acid will lead to lower band intensity in western blot. But with higher background band at the lower size. It seems for me there's some degradation of these samples. But i supposed the protein was prepared simultaneously and equally. And the experiment repeated twice with same results. Below are the attached pic.
Anyone knows why some sample will have higher intensity of unspecific band than the others?
Edited by Thomson, 27 August 2018 - 05:00 PM.