Disclosed are a pretreatment solution for immunohistochemical staining, which elutes a paraffin-containing embedding medium from a glass slide with a tissue specimen embedded in the medium, and retrieves antigenicity of the tissue specimen, and which is usable three or more times, and a pretreatment solution concentrate for immunohistochemical staining which allows ready preparation of the pretreatment solution and a method of immunohistochemical staining using the same. The pretreatment solution contains an antigen retrieving agent, particular nonionic surfactants, and cyclodextrin or a derivative thereof, with the balance being not less than 80 mass % of water. The content of the antigen retrieval agent is such that the pH of the pretreatment solution is in a predetermined range, and the content of cyclodextrin or a derivative thereof is a particular amount.