This report presents a guide to selecting high-quality commercial antibodies against Optineurin by immunoblot (Western blot), immunoprecipitation and immunofluorescence, using a knockout based validation approach. The research displayed in this study can be considered a subsequent study following the initial Optineurin report published to the YCharOS community in 2021 (DOI:10.5281/zenodo.4730992). Antibodies ab213556**, 60293-1-Ig*, 702766** and 711879**, previously characterized in the initial report, were retested in all three applications, employing our revised standardized protocols. Additionally, new recombinant antibodies were subject to characterization. This study was funded in part by Genome Québec's Genomics Integration Program, awarded to the research laboratory of Peter S. McPherson.This target was nominated by the ALS-Reproducible Antibody Platform (ALS-RAP), established as a public-private partnership by three prominent ALS charities - the ALS Association (USA), the Motor Neurone Disease Association (UK), and the ALS Society of Canada.