LARGE-SCALE AMINO-ACID-ANALYSIS FOR PROTEOME STUDIES

Amino-acid analysis is a relatively new method for identification of p roteins separated by two-dimensional gel electrophoresis and blotted o nto polyvinylidene difluoride (PVDF) membranes. This article describes modified amino-acid analysis methods for this purpose. Streamlined sa mple handling is a key feature of the process. To minimise sample mani pulation, a single vial is used for hydrolysis and the protein hydroly sate on PVDF membrane is extracted by a one-step procedure. The hydrol ysate should not be stored for long periods before analysis. Applicati ons of the technique are presented to demonstrate the identification p rocedure. This approach is the most cost-effective and time-effective first step in mass protein screening for a large-scale proteome projec t.