We have established a monoclonal antibody, GGR41, specific for a new class of minor gangliosides, such as GQ1b alpha and GT1a alpha, by immunizing mice with a GQ1b-rich ganglioside fraction extracted from bovine brain. Each of those minor gangliosides has been reported to be one of the cholinergic-specific gangliosides (Chol-1). Careful examination of binding specificity of the antibody by both an enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatograms showed that the antibody recognizes three sialyl residues separately attaching to the gangliotetraosyl backbone structure. Immunohistochemical analysis revealed that GGR41 immunostained lamina I and III of dorsal horn and lateral horn of human thoracic cord but motor neurons were not immunostained. Except for negative staining of motor neurons, this distribution is similar to the distribution pattern of staining as reported in rats and humans using a polyclonal antibody against Chol-1. Thus, the antibody obtained in this study should be a useful reagent to study the function of a unique new class of the minor gangliosides.