Murine monoclonal antibodies (MAbs) were produced which were specific for Pasteurella haemolytica serotype 1 lipopolysaccharide (LPS). The MAbs also reacted with LPS present in a partially purified antigen derived from a saline extract of the organism. The epitope to which the MAbs were directed was a carbohydrate which was sensitive to oxidation with periodate, had a molecular weight between 14,000 and 25,000 as determined by immunoblotting, and was present in a crude O-antigen preparation of P. haemolytica LPS. The MAbs did not react with purified capsular polysaccharide from P. haemolytica serotype 1. In an enzyme-linked immunosorbent assay, reaction of the MAbs with LPS obtained from 14 gram-negative bacteria failed to detect any cross-reactivity with P. haemolytica LPS. However, the MAbs detected antigenic similarities among P. haemolytica serotypes 1, 5, 6, 7, 8, and 12 and, to a lesser extent, 4 and 14. These studies indicate that the LPS-O-antigens from several P. haemolytica serotypes have similar epitopes and may be partially responsible for shared antigenicity among serotypes.