Does adding triton with the primary antibody hinder antigen detection in immunohistochemistry?

In general, it is best to use Triton X-100 prior to incubation with the primary antibody and exclude the Triton X-100 during primary antibody incubation. This is because Triton X-100 is a harsh detergent. If used during primary antibody incubation, it has the potential to remove epitopes from tissue, which will reduce the signal intensity.