We convened an ad hoc International Working Group for Antibody Validation in order to formulate the best approaches for validating antibodies used in common research applications and to provide guidelines that ensure antibody reproducibility. We recommend five conceptual 'pillars' for antibody validation to be used in an application-specific manner.

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Acknowledgements

The authors are responsible for all content and editorial decisions and received no honoraria related to the development of this publication. Editorial assistance in the preparation of this publication was provided by Phase Five Communications, supported by Thermo Fisher Scientific, which had no other involvement in the development of this publication.

Author information

Affiliations

  1. Science for Life Laboratory, School of Biotechnology, KTH Royal Institute of Technology, Stockholm, Sweden.

    • Mathias Uhlen
    •  & Emma Lundberg
  2. Center for Research in Biological Systems, University of California at San Diego, San Diego, California, USA.

    • Anita Bandrowski
  3. Proteomics Platform, Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, Massachusetts, USA.

    • Steven Carr
  4. Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.

    • Aled Edwards
  5. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

    • Jan Ellenberg
  6. Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, USA.

    • David L Rimm
  7. Office of Cancer Clinical Proteomics Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.

    • Henry Rodriguez
    •  & Tara Hiltke
  8. Department of Genetics, Stanford University School of Medicine, Stanford, California, USA.

    • Michael Snyder
  9. Biofluid Biomarker Center, Niigata University, Niigata, Japan.

    • Tadashi Yamamoto

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Competing interests

M.U. is cofounder of Atlas Antibodies, Affibody Medical, and Antibodypedia. S.C. is a consultant to Biogen and PTM BioLabs Inc. A.B. is the founder of SciCrunch Inc., the technology backing for the Resource Identification Initiative. E.L. acknowledges formal links to Atlas Antibodies. D.L.R. is a consultant to Amgen, Applied Cellular Diagnostics, AstraZeneca, Agendia, Bethyl Labs, Biocept, BMS, Cernostics, FivePrime, Genoptix/Novartis, Metamark Genetics, MDAgree, OptraScan, and Perkin Elmer; he has received honoraria from Genentech/Roche and Ventana; and he acknowledges research support from Cepheid, Genoptix, Gilead Sciences, Kolltan, Perkin Elmer, and Nantomics. D.L.R. holds equity in MDAgree.

Corresponding author

Correspondence to Mathias Uhlen.

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https://doi.org/10.1038/nmeth.3995