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Kinases belong to one of the most biologically§significant enzyme classes. The development of§analytical techniques for characterization of kinase§activity, in particular at a global scale, is a§central priority for proteomic and cell biology§researchers.§In order to facilitate global analysis of cellular§phosphorylation, a new paradigm of microarray§technology which focuses on analysis of total§cellular kinase activity, kinome, has emerged in the§past few years.§As the specificity of many kinases is dictated§primarily by recognition of residues immediately§surrounding the site of phosphorylation a logical§methodology is to employ peptides§representing these immediate sequences as§experimental substrates. Microarray chips carrying§hundreds of such substrate targets have been§developed for human kinome analysis, however,§lack of similar tools for species outside research§mainstream has limited kinome analysis in§these species. This work describes a methodology to§create species specific peptide arrays. A case study§is carried out to analyze Toll Like Receptor (TLR)§signaling pathways in bovine monocytes.