Generation and Evaluation of an Anti-REV1 Monoclonal Antibody

Continuous exposure of cells to exogenous and endogenous agents produces many types of DNA damage during normal cell cycles. Post-replication repair, consisting of error-free and error-prone sub-pathways, is required for tolerance of such DNA damage. REV1 plays a crucial role in regulation of the error-prone pathway. To facilitate analysis of its cellular functions, we here generated a mouse REV1 monoclonal antibody, called D6, which also recognizes human REV1. The epitope for the antibody could be mapped between 860-877 amino acid residues of human REV1, which was located outside of the conserved catalytic domain. Although the antibody unfortunately could not specifically detect endogenous mouse and human REV1 by western blotting and immunohistochemistry, we established a method to identify endogenous human REV1 by immunoprecipitation-western blotting analysis combining D6 and separately generated polyclonal antibodies.