I am trying to do GST pull down with my experimental proteins and GST as my negative control. however, my negative control (GST) is interacting very strongly to my target protein and even lowering the intensity on Li-cor odyssey it still shows as compare to my positive controls and my experimental (target/bait proteins). I have literally tried almost everything and even kept an eagle eye on my washing part but still to no avail. Can anyone please suggest how to keep negative control as negative. thanks
p.s: first lane on the left is the GST negative control followed by my experimental.