Abstract: The photoreceptor G protein transducin [α- and βγ-subunits (Tα/Tβγ)] plays a central role in the visual transduction process. The amino-terminus of bovine Tα is modified by one of four distinct fatty acids—laurate (C12:0), myristate (C14:0), C14:1 (5-cis), and C14:2 (5-cis, 8-cis)—but the biological significance and the localization of the four isoforms of Tα are poorly understood. To investigate the cellular distribution of each isoform, we prepared monoclonal antibodies against a synthetic C12:0-, C14:0-, C14:1-, or C14:2-nonapeptide corresponding to the N-terminal region of Tα. Among several types of antibodies isolated, only one type, represented by LA4, reacted specifically with the C12:0-peptide as well as purified Tα but not with the other proteins in bovine retinal homogenate, including recoverin, indicating that the epitope comprises both C12:0 and the N-terminal amino acids of Tα. Immunohistochemical analyses of bovine retinal sections by LA4 showed the uniform distribution of C12:0-Tα in almost all the rod outer segments. Hence, it seemed unlikely that each isoform of Tα was localized in specific cells. This observation, together with evidence for a possible functional diversity among the isoforms, suggests that the four isoforms of Tα in a single rod cell may contribute simultaneously to a fine tuning of the photon-signal transduction process.