Preparation of anti-hECSM2 mouse monoclonal antibodies and their application in the analysis of hECSM2 expression.

Human endothelial cell-specific molecule 2 (hECSM2) is a novel, recently identified gene, the biological functions of which are still unclear. The aim of this study was to prepare anti-hECSM2 mouse monoclonal antibodies and investigate the endogenous expression of hECSM2 in cell lines and human tissues. Mouse monoclonal antibodies (MAbs) specifically against hECSM2 were prepared using the hybridoma method. Western blot and flow cytometry were used to detect the specificity of the antibodies. Immunofluorescence and immunohistochemistry were used to investigate the endogenous expression of hECSM2 in different kinds of cell lines and human tissues, respectively. Two anti-hECSM2 MAbs secreting hybridomas were selected. Experiments showed that these two antibodies were highly specific to hECSM2 and endogenous hECSM2 was located on the endothelial cell membrane. Our anti-hECSM2 mouse antibodies can be used for Western blot, flow cytometry, and immunohistochemistry study, and can be a valuable tool for investigating the function and distribution of hECSM2.