Protein kinases (PKs) play crucial roles in intracellular signal transduction and they are one of the representative targets of drug development because their hyper-activation is related to many diseases including cancer. Kinomics is a technique to evaluate the activities of whole cellular PKs and will be important for drug development and diagnosis for personalized medicine. Peptide microarray is one of the representative methods to evaluate kinomics. We have proposed several techniques to improve the detection of peptide microarrays and have achieved quantitative evaluation of cellular PKs based on both fluorescence and SPR detection. We have also succeeded in screening of highly reactive substrate peptides of target PKs by using our microarrays.