Parvalbumin is the major allergen of fish species. Investigation into this protein is beneficial not only to detecting allergen, but also to producing aquatic products with low allergenicity. In the present study, parvalbumin from the skeletal muscle of silver carp (Hypophthalmichthy molitrix) was first purified to homogeneity by homogenization, centrifugation, heat treatment extraction and gel filtration chromatography on Superdex 75. Purified parvalbumin revealed three protein bands with molecular mass of 12, 14 and 24 ku, respectively, as detected by Tricine-SDS-PAGE under nonreducing conditions while under reducing conditions, only a band corresponding to 12 ku was detected. Westernblotting using antifrog parvalbumin monoclonal antibody (PARV-19) positively reacted with the three protein bands of 12, 14 and 24 ku, suggesting they are different forms of parvalbumin. A polyclonal rabbit antisilver carp parvalbumin antibody was prepared and immunoglobulin G (IgG) was purified by Protein A Sepharose affinity column. Dot-blot revealed that the antibody reacted with parvalbumin even after dilution to 1/51200, suggesting its higher titer. Westernblotting analysis indicated that parvalbumins from four fishes (common carp, silver carp, crucian carp, sea bream) can all be detected by the polyclonal antibody specifically.