Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia–pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti‐HER2 and anti‐Ki‐67. The aim of this study was to evaluate the expression of immunohistochemical markers HER‐2 and Ki‐67 in granulosa cell tumour from bitches´ ovaries. In HER‐2 immunomarker analysis using the HercepTest® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki‐67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti‐HER‐2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti‐Ki‐67 and anti‐HER‐2 antibody immunostaining intensities, probably due to only one case with a high Ki‐67 index. With an effective protocol for HER‐2 and Ki‐67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.