Four rabbit antibodies have been prepared, which are specifically directed against alpha-lactalbumins from different sources; namely human, cow, goat and sheep milk. Each of these antibodies was tested for its ability to react with, separately, each of the four proteins. The immunological reactions were assessed by means of different techniques: double immunodiffusion in agar gel as well as affinity chromatography of antibodies, using antigens covalently bound to an insoluble matrix. In each case, the strongest reaction was observed between homologous antibody and (matrix-bound) antigen; heterologous antigens were, however, also capable of cross-reaction. Whereas no cross-reaction between human alpha-lactalbumin and antibodies against the bovine protein could be evidenced by immunodiffusion, the occurrence of soluble complexes has been disclosed by means of a gel filtration technique.