Klugmann et al. Rare Dis Orphan Drugs J 2023;2:8 DOI: 10.20517/rdodj.2023.05 Rare Disease and Orphan Drugs Journal © The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, sharing, adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. www.rdodjournal.com Open Access Technical Note Histological and biochemical methods to assess aminoacyl-tRNA synthetase expression in human post-mortem brain tissue Matthias Klugmann, Alexandra K. Suchowerska, Gary D. Housley, Dominik Fröhlich Translational Neuroscience Facility & Department of Physiology, School of Biomedical Sciences, UNSW Sydney, Sydney NSW 2052, Australia. Correspondence to: Dominik Fröhlich, MSc, PhD, Translational Neuroscience Facility & Department of Physiology, School of Biomedical Sciences, UNSW Sydney, Sydney NSW 2052, Australia. E-mail: d.frohlich@unsw.edu.au How to cite this article: Klugmann M, Suchowerska AK, Housley GD, Fröhlich D. Histological and biochemical methods to assess aminoacyl-tRNA synthetase expression in human post-mortem brain tissue. Rare Dis Orphan Drugs J 2023;2:8. https://dx.doi.org/10.20517/rdodj.2023.05 Received: 28 Jan 2023 First Decision: 2 Mar 2023 Revised: 15 Mar 2023 Accepted: 10 Apr 2023 Published: 20 Apr 2023 Academic Editors: Daniel SCHERMAN, Jacques S Beckmann Copy Editor: Ying Han Production Editor: Ying Han Abstract Aminoacyl-tRNA synthetases are essential, non-redundant enzymes that catalyze the charging of tRNAs with their cognate amino acids. This reaction is a prerequisite for protein translation in all cells. Mutations in human aminoacyl-tRNA synthetases are often associated with defects of the peripheral and central nervous system and are the underlying cause of many rare diseases including neuropathies and leukodystrophies. A comprehensive understanding of aminoacyl-tRNA synthetase expression domains is key to understanding these disorders and developing novel targeted treatment strategies. Here, we describe histological and biochemical methods to analyze the expression pattern of the aspartyl-tRNA synthetase AspRS in human post-mortem brain tissue. The same methods can readily be applied to other members of the aminoacyl-tRNA synthetase superfamily or, more generally, to other cytosolic proteins in the human brain. Keywords: Aminoacyl-tRNA synthetase, aspartyl-tRNA synthetase, AspRS, DARS1, post-mortem brain tissue INTRODUCTION Protein synthesis is an evolutionarily highly conserved process that requires an intimate interplay of