The identification of high-performing antibodies for Sequestosome-1 for use in Western blot, immunoprecipitation and immunofluorescence

• DOI: 10.12688/f1000research.132628.1
• Publication Date: 2023-03-23T16:45:07Z
• Authors: Riham Ayoubi; Walaa Alshafie; Irina Shlaifer; Kathleen Southern; Peter S. McPherson; Carl Laflamme
• Keywords: Sequestosome-1 Protein; Blotting, Western; Humans; Fluorescent Antibody Technique; Immunoprecipitation; Data Note; Antibodies

Sequestosome-1, encoded by the gene SQSTM1, functions as a bridge between ubiquitinated proteins and the proteasome or autophagosome, thereby regulating protein degradation pathways. Loss of Sequestosome-1 is hypothesized to enhance neurodegeneration progression in several diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal disorders (FTD). Sequestosome-1 reproducible research would be facilitated with the availability of well-characterized anti-Sequestosome-1 antibodies. In this study, we characterized seventeen Sequestosome-1 commercial antibodies for Western blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.