Immunofluorescence (IF/ICC) Troubleshooting: Why the immune fluorescence result has non-specific staining?

If your immunefluorescence results have non-specific staining, the main possible reasons are as follows.
1. You should make confirm that the section or cells have dried out. In all process, the sections or cells should be at high humidity and do not let them dry out.
2. The antibody concentration is too high. The antibody includes primary antibody and second antibody. Use lower concentration of antibody in the next test.
3. In the process of immunefluorescence, the enzyme inhibitors can inactive endogenous peroxidases.
4. The primary antibody should raised in species which is different from the cell or tissue. If your tissue or cells are from mouse, primary antibody is should not mouse antibody and should be antibody from different species.