Abstract: There are 4 tyrosine residues in succinyl CoA: 3-oxoacid CoA transferase (SCOT), in which nitration of Tyr4 and Tyr76 will lead to change of the structure and decrease of enzymatic activity. In this study artificial synthetic peptides covering the SCOT Tyr4 and Tyr76 region with the tyrosine nitrated were employed as hapten and conjugated to keyhole limpet hemocyanin (KLH) to immunize Balb/c mouse. The monoclonal antibodies against nitrated Tyr4 and Tyr76 in SCOT are developed. After screening of massive cell lines, 4 antibodies against nitrated Tyr4 and tyr76 epitope of SCOT respectively were selected and determined by indirect ELISA. The result showed that the titer of the antibodies were 1∶128 000, 1∶32 000, 1∶512 000 and 1∶4 096 000 respectively. The results indicated that the antibodies obtained showed good specificity and sensitivity which will lay a foundation for further study of SCOT nitrotyosine modification and its impact on physiology. .