Journal article
A consensus platform for antibody characterization
Nature protocols, Vol.20(6), pp.1509-1545
06/2025
DOI: 10.1038/s41596-024-01095-8
PMCID: PMC13054593
PMID: 39690206
Abstract
Antibody-based research applications are critical for biological discovery. Yet there are no industry standards for comparing the performance of antibodies in various applications. We describe a knockout cell line-based antibody characterization platform, developed and approved jointly by industry and academic researchers, that enables the systematic comparison of antibody performance in western blot, immunoprecipitation and immunofluorescence. The scalable protocols, which require minimal technological resources, consist of (1) the identification of appropriate cell lines for antibody characterization studies, (2) development/contribution of isogenic knockout controls, and (3) a series of antibody characterization procedures focused on the most common applications of antibodies in research. We provide examples of expected outcomes to guide antibody users in evaluating antibody performance. Central to our approach is advocating for transparent and open data sharing, enabling a community effort to identify specific antibodies for all human proteins. Mid-level graduate students with training in biochemistry and prior experience in cell culture and microscopy can complete the protocols for a specific protein within 1 month while working part-time on this effort. Antibody characterization is needed to meet standards for resource validation and data reproducibility, which are increasingly required by journals and funding agencies.
Details
- Title: Subtitle
- A consensus platform for antibody characterization
- Creators
- Riham Ayoubi - Montreal Neurological Institute and HospitalJoel Ryan - McGill UniversitySara Gonzalez Bolivar - McGill UniversityCharles Alende - Montreal Neurological Institute and HospitalVera Ruiz Moleon - McGill UniversityMaryam Fotouhi - Montreal Neurological Institute and HospitalMona Alqazzaz - University of TorontoKathleen Southern - Montreal Neurological Institute and HospitalWalaa Alshafie - Montreal Neurological Institute and HospitalMatt R Baker - Thermo Fisher Scientific (United States)Alexander R Ball Jr - GeneTex (United States)Danielle Callahan - Proteintech Group, Rosemont, IL, USAJeffery A Cooper - R&D Systems (United States)Katherine Crosby - Cell Signaling Technology (United States)Kevin J Harvey - Aviva Systems Biology, San Diego, CA, USADouglas W Houston - University of IowaRavindran Kumaran - Abcam (United Kingdom)Meghan Rego - AddgeneChristine Schofield - Revvity, Waterbeach, Cambridge, UKHai Wu - ABclonal Technology, Woburn, MA, USAMichael S Biddle - NIHR Leicester Biomedical Research CentreClaire M Brown - McGill UniversityRichard A Kahn - Emory UniversityAnita Bandrowski - University of California San DiegoHarvinder S Virk - University of LeicesterAled M Edwards - Structural Genomics ConsortiumPeter S McPherson - Montreal Neurological Institute and HospitalCarl Laflamme - Montreal Neurological Institute and Hospital
- Resource Type
- Journal article
- Publication Details
- Nature protocols, Vol.20(6), pp.1509-1545
- DOI
- 10.1038/s41596-024-01095-8
- PMID
- 39690206
- PMCID
- PMC13054593
- NLM abbreviation
- Nat Protoc
- ISSN
- 1750-2799
- eISSN
- 1750-2799
- Publisher
- NATURE PORTFOLIO; BERLIN
- Grant note
- OGI-210 / Genome Canada (Génome Canada) 18331 / Michael J. Fox Foundation for Parkinson's Research (Michael J. Fox Foundation) RF1AG057443 / U.S. Department of Health & Human Services | NIH | National Institute on Aging (U.S. National Institute on Aging)
- Language
- English
- Electronic publication date
- 12/17/2024
- Date published
- 06/2025
- Academic Unit
- Biology
- Record Identifier
- 9984758187702771
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