Ovine polyclonal antisera were raised to aminolevamisole to develop an enzyme immunoassay for levamisole. The assay has been validated in milk and in 100 g l^–1 muscle homogenate. The limit of detection is 1 µg l^–1. In milk the inter-assay relative standard deviation (s_r) is 6% at 10 µg kg^–1 and the intra-assay s_r value 7.2%. In tissue homogenate the inter-assay s_r value is 12.7% at 5 µg kg^–1 and the intra-assay s_r value is 3.8%. Total incubation time is 1 h.