A validated antibody toolbox for ALS research
11 affiliations
- 1Structural Genomics Consortium, Montreal Neurological Institute-Hospital (The Neuro), McGill University, Montreal, Canada
- 2Neurodegenerative Diseases Research Group, Department of Neurology and Neurosurgery, Montreal Neurological Institute-Hospital (The Neuro), McGill University, Montreal, Canada
- 3NIHR Respiratory Biomedical Research Centre, Department of Respiratory Sciences, University of Leicester, Leicester, United Kingdom
- 4Early Drug Discovery Unit (EDDU), Department of Neurology and Neurosurgery, Montreal Neurological Institute-Hospital (The Neuro), McGill University, Montreal, Canada
- 5Department of Genetics, Stanford University School of Medicine, Stanford, United States
- 6Structural Genomics Consortium, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
- 7Centre for Human Genetics and Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
- 8Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
- 9Structural Genomics Consortium, Department of Medicine, Karolinska Institute, Solna, Sweden
- 10Neuroimmunology Unit, Department of Neurology and Neurosurgery, Montreal Neurological Institute-Hospital (The Neuro), McGill University, Montreal, Canada
- 11School of Pharmaceutical Sciences, Faculty of Medicine, University of Ottawa, Ottawa, Canada
Abstract
A substantial fraction of amyotrophic lateral sclerosis (ALS)-associated proteins remains poorly characterized, in part because of the limited availability of validated research antibodies. We established knockout (KO)-based antibody characterization workflows and demonstrated that widely used antibodies against the major ALS-associated protein C9orf72 lacked specificity (Laflamme et al., 2019). We subsequently scaled this framework to systematically benchmark research antibodies, revealing that up to 61% fail to perform as recommended by manufacturers (Ayoubi et al., 2023b). Here, we extend this approach by establishing the ALS-Reproducible Antibody Platform (ALS-RAP), a comprehensive effort to generate a publicly available dataset of KO-validated antibodies targeting proteins encoded by ALS risk genes. In total, we characterized 303 antibodies against 33 ALS-associated proteins to identify high-quality reagents for use in western blot, immunoprecipitation, and immunofluorescence. Using these antibodies, we profiled protein levels across human induced pluripotent stem cell-derived and primary neurological cell types. These analyses revealed diverse cellular distributions and higher levels of several ALS-associated proteins in glial populations, consistent with emerging evidence for immune contributions to ALS. Together, ALS-RAP provides a validated antibody toolbox and protein expression resource to support the study of ALS-associated proteins.
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BibTeX
@article{ayoubi2026validated,
title = {{A validated antibody toolbox for ALS research}},
author = {Riham Ayoubi and Emma J MacDougall and Ian McDowell and Michael S Biddle and Bárbara T Ferreira and CongYao Zha and Marie-France Dorion and Jay P Ross and Sara Gonzalez Bolivar and Vera Ruiz Moleon and Charles Alende and Vincent Francis and Maryam Fotouhi and Mathilde Chaineau and Carol X Chen and Valerio EC Piscopo and Vincent Soubannier and Irina Shlaifer and Zhipeng You and Michael Nicouleau and Tracy Keates and Wen Hwa Lee and Brian D Marsden and Leonidas Koukouflis and Edvard Wigren and Carolyn A Marks and Luke M Healy and Patrick A Dion and Guy A Rouleau and Edward A Fon and Harvinder S Virk and Susanne Gräslund and Opher Gileadi and Aled M Edwards and Thomas M Durcan and Peter S McPherson and Carl Laflamme},
journal = {eLife},
year = {2026},
month = nov,
doi = {10.7554/elife.111349},
url = {https://elifesciences.org/articles/111349},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - A validated antibody toolbox for ALS research
AU - Riham Ayoubi
AU - Emma J MacDougall
AU - Ian McDowell
AU - Michael S Biddle
AU - Bárbara T Ferreira
AU - CongYao Zha
AU - Marie-France Dorion
AU - Jay P Ross
AU - Sara Gonzalez Bolivar
AU - Vera Ruiz Moleon
AU - Charles Alende
AU - Vincent Francis
AU - Maryam Fotouhi
AU - Mathilde Chaineau
AU - Carol X Chen
AU - Valerio EC Piscopo
AU - Vincent Soubannier
AU - Irina Shlaifer
AU - Zhipeng You
AU - Michael Nicouleau
AU - Tracy Keates
AU - Wen Hwa Lee
AU - Brian D Marsden
AU - Leonidas Koukouflis
AU - Edvard Wigren
AU - Carolyn A Marks
AU - Luke M Healy
AU - Patrick A Dion
AU - Guy A Rouleau
AU - Edward A Fon
AU - Harvinder S Virk
AU - Susanne Gräslund
AU - Opher Gileadi
AU - Aled M Edwards
AU - Thomas M Durcan
AU - Peter S McPherson
AU - Carl Laflamme
PY - 2026
DA - 2026/11/03
JO - eLife
DO - 10.7554/elife.111349
UR - https://elifesciences.org/articles/111349
AB - A substantial fraction of amyotrophic lateral sclerosis (ALS)-associated proteins remains poorly characterized, in part because of the limited availability of validated research antibodies. We established knockout (KO)-based antibody characterization workflows and demonstrated that widely used antibodies against the major ALS-associated protein C9orf72 lacked specificity (Laflamme et al., 2019). We subsequently scaled this framework to systematically benchmark research antibodies, revealing that up to 61% fail to perform as recommended by manufacturers (Ayoubi et al., 2023b). Here, we extend this approach by establishing the ALS-Reproducible Antibody Platform (ALS-RAP), a comprehensive effort to generate a publicly available dataset of KO-validated antibodies targeting proteins encoded by ALS risk genes. In total, we characterized 303 antibodies against 33 ALS-associated proteins to identify high-quality reagents for use in western blot, immunoprecipitation, and immunofluorescence. Using these antibodies, we profiled protein levels across human induced pluripotent stem cell-derived and primary neurological cell types. These analyses revealed diverse cellular distributions and higher levels of several ALS-associated proteins in glial populations, consistent with emerging evidence for immune contributions to ALS. Together, ALS-RAP provides a validated antibody toolbox and protein expression resource to support the study of ALS-associated proteins.
SN - 2050-084X
ER -