Monoclonal antibodies to a brain dopamine binding protein: Production, specificity, and immunohistochemistry

Abstract

A dopamine binding protein (DABP) has been purified from the rat brain synaptic membrane to homogeneity by affinity chromatography and gel electrophoresis. The monoclonal antibodies against the DABP were produced by the mouse-mouse hybridoma technique and characterized for their specificity to dopamine receptors by displacement of dopamine receptor binding. These monoclonal antibodies have been used to localize DABP in rat brain by immunohistochemistry. A specific linear structure of reaction product was seen in both caudate nucleus and cerebral cortex. This finding suggests that the DABP is present in the cerebral cortex and caudate nucleus as a membranous component of the neurons or their processes.

Antibodies Against Synthetic Peptides Predicted from the Nucleotide Sequence of D2 Receptor Recognize Native Dopamine Receptor Protein in Rat Striatum

“…Attempts by previous investigators to produce DA receptor antibodies were aimed via antiidiotypic routes, employing antibodies specific for DAergic drugs (Abbot and Strange, 1986). Hsu et al (1987) kDa DA binding protein and speculated the protein to be a subunit of DA receptor, but the presence of such a subunit structure in the DA receptor has not been confirmed. Several lines of experimental evidence support the notion that the peptide I antibody presented in this article is directed against the DA D2 receptor.…”