CD8 molecule is a key marker on T cell surface and is connected with the antigen recognition and activation of T lymphocytes. In order to provide a detection method for quantifying goose CD8 alpha expression, this study raised the protein and antibody for goose CD8 alpha and developed a feasible cell marker enzyme-linked immunoabsorbent assay (ELISA) method. Recombinant protein of the extracellular region gene of goCD8 alpha was expressed in prokaryotic expression system, and specific polyclonal antibodies for goCD8 alpha were raised and purified, which was further confirmed by Western-blot, immunofluorescence assay (IFA), and immunohistochemistry (IHC). A cell marker ELISA was established and optimized to detect the change of goCD8 alpha expression between goose parvovirus (GPV)-infected and mock-infected goose peripheral blood mononuclear cells (PBMCs), which is consistent with our previously results of real-time quantitative PCR (qPCR). Cell marker ELISA can provide a new method to detect goCD8 alpha in protein level and in a sensitive, specific, and simple way. This may provide a convenient and novel method for the detection of goCD8 alpha expression.