Previous immunostaining protocols are highly specific for model organisms and often not suitable for diverse specimens that are non-perfused and over-fixed (i.e., tissues sitting in fixatives for months/year). Here, we present an immunofluorescence protocol for localizing protein targets in brain tissue from 11 model and non-model mammals. We describe preparation of both fresh and fixed tissues including steps for deparaffinization, fixation, and cryoprotection.
