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The present study was focused on investigating the acute toxicity of chlorpyrifos and its impact on electrophoretic protein patterns in the tissues of Cyprinus carpio, an acute toxicity bioassay was conducted in the semi-static system and the fingerlings of C. carpio were exposed to various chlorpyrifos(50%EC)concentrations ranging from 0.1mg/L to 0.7mg/L for 96 h, and acute toxicity was determined as 0.318mg/L. Based on the LC50 value, the Cyprinus carpio were exposed to two sub-lethal concentrations viz., SL-1 (1/5th of LC50, 0.0636mg/l) and SL-2 (1/10th of LC50,0.0318 mg/l) of chlorpyrifos. Proteins have a unique role in cell metabolism and proteomic analysis provides valuable information when variations that occur within the proteome of organisms are compared as a consequence of biological perturbations or external stimuli. The study's goal was to investigate the influence of chlorpyrifos on electrophoretic protein patterns by performing SDS-PAGE on muscle, liver, gill, and brain tissues after exposing fish to two sublethal concentrations in a semi-static system for 3 weeks. The intensity of protein subunits in exposed tissues decreased, and some subunits disappeared, CPF influences both high-intensity protein bands and low intensity as well in both the sub-lethal concentrations. The study has suggested that the changes in the protein banding are more prominent in the SL-1 concentration of chlorpyrifos.The variations in protein subunits band patterns are due to protein deprivation of various proteins, proteolysis, or maybe insufficient amino acid incorporation into the polypeptide chain due to induced stress imposed by the toxicant.