A simple and rapid dot-immunobinding assay for zein and other prolamins

A simple method for the immunochemical assay of zein and other prolamins is described. Alcohol-solubilized zein is spotted onto chromatography paper disks or strips, incubated with antiserum (30-60 min), washed with several changes of Tris-buffered saline (10 min), incubated with protein A-peroxidase (30 min), and washed with several changes of Tris-buffered saline (10 min). the binding of zein-specific antibody is visualized by peroxidase-catalyzed color production from 4-chloro-1-naphthol. The method is simple, inexpensive, and rapid (2 h total time) and has potential application to other insoluble antigens.