This work demonstrates that Brucella lipopolysaccharide (LPS) preparations are a family of related molecules which display heterogeneity not only at the level of the O polysaccharide, but also at the core oligosaccharide and the lipid A. Sodium dodecyl sulphate/polyacrylamide gel electrophoresis and Western blotting showed that LPS from Brucella strains displayed molecular weight and antigenic heterogeneity. Smooth-type LPS (S-LPS) from B. abortus demonstrated three broad high-molecular-weight bands corresponding to S-LPS, and a low-molecular-weight band corresponding to O antigen lacking rough-type LPS (R-LPS). B. abortus R-LPS displayed four bands in increasing proportions as the molecular weight diminished. Immunodetection on high-performance thin-layer chromatography (HPTLC) with monoclonal antibodies (mAb) showed that R-LPS displayed three diffuse bands. HPTLC of O polysaccharide revealed two fast migrating bands recognized by antibodies. Gel chromatography and HPTLC analysis of core oligosaccharides from R-LPS demonstrated molecular weight heterogeneity as well as heterogeneous banding pattern, with no obvious correspondence between the two profiles. Immunodetection of lipid A on HPTLC plates revealed two major and three minor bands. Reactivity with mAbs suggested that regardless of the lipid A heterogeneity the basic structure of lipid A backbone is maintained.