Different protein fractionation techniques were used to define differences between a set of 8 wheat lines used in genetic mapping studies in Australia. A proteomics approach was used to establish the feasibility of identifying new protein polymorphisms for mapping purposes. Detailed analysis confirmed differences in the glutenin subunits, gliadin proteins, and 10-20 other proteins, between the mapping population parents, Cranbrook, Halberd, CD87, and Katepwa. Differences were particularly evident in the low molecular weight classes of protein. Alternative technologies were used to determine the differences in various protein classes in order to screen doubled haploid lines derived from crosses between the wheat lines. Polyacrylamide gel electrophoresis analysis allowed the mapping of loci encoding high molecular weight (HMW) and low molecular weight (LMW) glutenin subunit proteins. Reversed phase high performance liquid chromatography also allowed several loci encoding LMW glutenin subunit proteins to be mapped, as well as a new protein on chromosome 6A. Capillary electrophoresis provided a high-resolution system that was used to map several gliadin-type proteins. The studies showed that proteins provide useful genetic markers and the data are discussed from the point of view of the advantages that protein-based markers offer in providing both genotypic and phenotypic data.11 page(s