Genome sequencing of many model systems, including the human, yeast, Arabidopsis, and rice genomes, has been completed. As a complement to genomic techniques, proteomics has emerged to study the composition, structure, function, and interaction of the expressed proteins or proteome in a given cell, tissue, or organism. During the past several years, tremendous progress has been made with proteomic techniques in human and yeast, but progress has been slower in plants due to several factors. Proteomic techniques have been used in plants, especially in agriculturally important crops and weeds, to understand mechanisms of herbicide tolerance and weed resistance. Proteomic techniques, starting from protein extraction to protein identification, have been developed and advanced for large-scale proteomics studies, but limitations and problems for these techniques still exist for plant proteomics. Further technological improvements are needed to enhance quantitative and comparative large-scale proteomics studies. Applications of these techniques may help weed scientists to understand stress tolerance in crops and investigate weediness traits further. Proteomic techniques have unique strengths but also offer several challenges. Together with transcriptomics and metabolomics, these techniques for analyzing global patterns of gene expression offer new and novel techniques for better understanding biological questions of interest to weed scientists in the future.