Anti-RBM3 polyclonal antibody

Although it has been described and generally accepted that the myocardium does not show any ischemic types of changes or other lesions in fatal carbon monoxide poisoning victims, we previously observed detailed pathological findings with careful staining using HE and/or Azan. We have used immnohistochemistry on the myocardium from twenty victims of both carbon monoxide poisoning and house fire. We especially selected antibodies which were previously recognized as cold stress antigens. And we also used antibodies related to stress, apoptosis, and vascular genesis. The nuclei showed good reactivity with CIRBP, RBM3, and SIRT1 antibodies. Although CIRBP and RBM3 were previously recognized as cold stress antigens, this study indicated that these antigens were also transcribed in hypoxia. Antigens related to stress, apoptosis, and vascular genesis were also detected in the myocardium from carbon monoxide poisoning victims. The results obtained in the present study suggest that antigens, such as CIRBP, RBM3 and SIRT1, may be useful temporal markers of global hypoxia or ischemia in myocardium. This study also found that the cells once exposed to stress and hypoxia, develop and activate signal pathways to prevent harmful consequences, and the expressed antigens may be retained after cell death and during the postmortem interval.

Purpose: In this study, we investigated the prognostic impact of human RBM3 expression in colorectal cancer using tissue microarray-based immunohistochemical analysis. Experimental design: One polyclonal antibody and four monoclonal anti-RBM3 antibodies were generated and epitope mapped using two different methods. Bacterial display revealed five distinct epitopes for the polydonal antibody, while the four mouse monoclonal antibodies were found to bind to three of the five epitopes. A peptide suspension bead array assay confirmed the five epitopes of the polydonal antibody, while only one of the monoclonal antibodies could be mapped using this approach. Antibody specificity was confirmed by Western blotting and immunohistochemistry, including siRNA-mediated knock-down. Two of the antibodies (polydonal and monoclonal) were subsequently used to analyze RBM3 expression in tumor samples from two independent colorectal cancer cohorts, one consecutive cohort (n = 270) and one prospectively collected cohort of patients with cancer of the sigmoid colon (n = 305). RBM3-expression was detected, with high correlation between both antibodies (R = 0.81, p < 0.001). Results: In both cohorts, tumors with high nuclear RBM3 staining had significantly prolonged the overall survival. This was also confirmed in multivariate analysis, adjusted for established prognostic factors. Conclusion and clinical relevance: These data demonstrate that high tumor-specific nuclear expression of RBM3 is an independent predictor of good prognosis in colorectal cancer.