since the end of last year, western blotting is giving me a hard time. After (first time) antibody incubation of my western blot I always get a weak signal of my wanted protein band. However after stripping my blot and incubation with the same protocol and antibodies I get a stronger signal.
Before (mid of last year and earlier) I did not have this problem. I did not change my protocol, checked everything and just don't know why it is this way.
Does somebody have the same problem or knows what the problem is?
Thanks a lot in advance.