C/EBP [CCAAT/enhancer-binding protein]-homologous protein gene (chop) which plays an important role in endoplasmic reticulum stress-induced apoptosis, was investigated here by RACE and qPCR in an aquaculture animal for the first time. The full-length cDNA sequences of loach (Misgurnus anguillicaudatus) chop were 2533bp, encoding 266 amino acids. chop revealed obvious changes in transcriptions during different early life stages of the loach, and the highest appeared at the 8-cell stage. Among different tissues, loach chop predominantly expressed in gill, spleen and gonad. Then, we performed a hydrogen peroxide (H2O2, a common-used disinfectant) stress trial to explore the role of loach chop, and three different concentrations (0 µM, 50 µM and 100 µM) of H2O2 were set. After the stress, half of the loaches from 100 µM group were dead, whereas all loaches from the other two groups were alive. Meanwhile, the activities of CAT, SOD and GPX decreased obviously with the extension of stress time and increased H2O2 concentration. The expression levels of gill chop in loaches from 100 µM group were significantly higher than those from the other two groups at the 12th hour to 24th hour. atf4 and bax, two proapoptotic genes, significantly up-regulated in 100 µM group compared with the other two groups at the 18th hour and 24th hour. While the expression profiles of an antiapoptotic gene bcl2 basically presented an opposite trend. These results indicated a close relationship between H2O2 stress and fish apoptosis, and loach chop played an important role in response to the stress.