A group of 14 monoclonal antibodies (mAbs) to staphylococcal enterotoxin B (SEB) were obtained by fusion of Sp2/O myeloma cells with spleen cells from female BALB/c mice immunized with commercial SEB. The antibodies belonged to IgG1 and IgG2b subclasses. We evaluated the anti-SEB titres, competition assays and sensitivity of detection by indirect ELISA. Reactivity and cross-reactivity were also studied by indirect ELISA and confirmed by immunoblotting. All the mAbs reacted with SEB and with a second band which had a different electrophoretic mobility and probably represents an aggregate of SEB or SEB bound to membranes. Three mAbs reacted only with SEB and the rest showed cross-reactions with SEC1. No reactions were observed against any other serovar (SEA, SED and SEE) or other proteins.