Immunohistochemistry (IHC) Tips
Incubation with primary and secondary antibodies:
The conditions in which samples are incubated with primary antibodies are vital. The factors that affect antibody incubation include incubation time, temperature and antibody concentration. Typically, 1-2h at 37°C or overnight at 4°C following 45 min at 37°C (recommended) is appropriate.
As for the incubation with secondary antibodies, 30min-1h at room temperature or 37°C is appropriate.