I performed my second ever Western blot and I am somewhat confused about my results. I was checking for the expression of a fungal effector protein in barley leaf extracts. In the short exposure, I saw much less (understandably) but the curious thing was that some neighbouring bands, of similar intensity, were not observed in the short exposure but were seen in the long exposure.
Short exposure (3 minutes):
Long exposure (30 minutes):
Any reason why this could be so?
Also, I loaded twice as much protein than in my first Western and I observed much more non-specific binding. Is this normal?