Understanding molecular interactions of bacteria with their environment requires the purification and characterization of cell wall components. Here, we describe detailed experimental methods for the extraction, purification, and analysis of wall teichoic acids (WTA), which assume important roles as major constituents of Gram-positive cell walls, such as mediating interaction with cell wall-associated proteins, eukaryotic host cells, and bacteriophages. Specifically, we present a procedure for compositional WTA characterization to study large diversity of carbohydrate substitution on Listeria monocytogenes WTA. This protocol may also be used and adapted to analyze WTA from other bacteria.