Insulin-like growth factor binding proteins produced by cultured rat neurons, astrocytes, and rat cell lines BRL-3A and B104 were compared to binding proteins found in rat serum, using affinity labeling, deglycosylation, and Western ligand blotting studies. Each source elaborated an unique pattern of heterogeneous binding proteins. Some of the binding proteins from different sources behaved similarly in each experimental system suggesting that subsets of these binding proteins may be structurally related. In particular, our data suggest that cultured astrocytes and neurons make the major binding protein produced by BRL-3A cells.