Antiphosphotyrosine antibodies for studying protein phosphorylation in neural cells: Applications to brain slices and cultured cells

Abstract

Antibodies against phosphotyrosine are a powerful tool for studying protein phosphorylation In intact cells. These antibodies can be used for immunoblotting, immunocytochemistry, immunoprecipitation, and affinity purification experiments. In this article we describe experimental protocols for preparing and purifying antiphosphotyrosine antibodies and for using them to study protein phosphorylation in intact brain, in brain slice preparations, and in nerve cell cultures. We also review some of the information that this approach has yielded on the development, characterization, and regulation of protein tyrosine phosphorylation in neurons and astrocytes.