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This paper reports an integrated online microdevice for protein analysis we developed. Our developed microdevice includes all steps of cell culture, pretreatment procedures and electrophoresis separation. In conventional method, pipetting and centrifugation are indispensable for washing cells, changing media and extraction of proteins during protein assay. Our developed system requires no external machine, such as centrifuge or pump. Our system can be free from troublesome pipetting. Utilizing the integrated online microdevice, dramatically reduced total analytical time (1 min per 12 samples) was achieved.