The specificity of antibodies to actin was assayed by use of immunoblots and histological sections of control tissues enriched for each of six different isoforms. On immunoblots, all antibodies stained at most one band of protein in most of the control materials, with a molecular weight of approximately 43 kDa. Their pattern of staining of muscle and nonmuscle tissues indicated their isoform specificity. On tissue sections, immunocytochemical staining demonstrated cellular and subcellular localization of the different isoforms. Once characterized with regard to specificity, these antibodies were used to probe actin in the guinea pig organ of Corti. None of the four muscle isoforms of actin were found in either immunoblots or tissue sections of the organ of Corti. Both beta- and gamma-cytoplasmic isoforms of actin were present in hair cells and supporting cells. This leaves open to investigation the role which cytoplasmic actins play in these cells of the organ of Corti.