I have had a problem with non-specific staining of vasculature in mouse brain IHC, particularly the cortex area. Somehow, it appears that the problem is from the secondary antibodies. Without the primary Abs, the secondary Abs with either fluorescence or DAB coloring still give the non-specific staining. Does anyone experience the same problem and know why?
If you help to solve this problem and give some tips, I would appreciate it.
Thanks,
Woongjai