I don't think I've not seen a thread on here about this yet, but if there is, I'm sorry for reposting
My initial probing of the Western Blot appears to go well, I can see that I've gotten pretty good transfer and can see my bands of interest.
When I strip the blot and go to reprobe it, I am not seeing anything beyond background banding. This includes not being able to visualize the ladder.
My thoughts were that I was too aggressive in my stripping and that I wasn't removing enough of the SDS or that my concentrations were too high. But I've double checked it all, have done excessive washes with TONS of TBST followed by TBS before reblocking and I still have this problem.
When I change species (my first blot is rabbit and after strip and reprobe the next set of antibodies is mouse) I get no signal at all, not even background levels.
Any help would be appreciated
Thanks in advance!