A new monoclonal antibody that recognizes alpha-actinin in cardiac muscle cells was used in a quantitative study (fluorescence activated cell sorting and polyacrylamide gel electrophoresis) of the expression of this protein during chick embryo development, to determine the changes in cytoplasmic and cytoskeletal compartments. alpha-Actinin expression was weak in early stages of development (Hamburger and Hamilton stage 18) and increased steadily until Hamburger Hamilton stage 40. In all stages, the protein was more abundant in the cytoplasmic compartment. The monoclonal antibody cross-reacted with alpha-actinin in chicken smooth and striated muscle cells and also showed a faint cross-reaction with human cardiac muscle alpha-actinin.