Objective To explore the effects of different methods of antigen retrieval on the results of immunohistochemical staining of nuclear receptors. Methods Antigens were retrieved by using microwave oven, autoclave, enzyme and autoclave plus enzyme, respectively, in liver sections from DA.1U rats, followed by immunohistochemical staining. Results After antigen retrieval with autoclave, the positive staining intensity of nuclear receptors LXR-α, LXR-β, PPAR-γ and FXR in the liver sections from DA.1U rats was enhanced obviously, and the location of nuclear receptors was better by using this method than the others. Conclusion In the immunohistochemical staining of nuclear receptors, using autoclave to retrieve antigens is the best method.