This antibody validation report describes an immunofluorescence analysis of an anti-NBR1 monoclonal antibody in rat spleen tissue. The tissue was processed using antigen retrieval, blocking, and staining with the primary and secondary antibodies overnight and 50 minutes respectively. It was then counterstained with DAPI and visualized under a fluorescence microscope. The report provides details of the immunofluorescence protocol including tissue processing, antigen retrieval, blocking, antibody incubations, mounting and visualization.