Guidelines on antibody use in physiology research

Am J Physiol Renal Physiol. 2024 Mar 1;326(3):F511-F533. doi: 10.1152/ajprenal.00347.2023. Epub 2024 Jan 18.

Abstract

Antibodies are one of the most used reagents in scientific laboratories and are critical components for a multitude of experiments in physiology research. Over the past decade, concerns about many biological methods, including those that use antibodies, have arisen as several laboratories were unable to reproduce the scientific data obtained in other laboratories. The lack of reproducibility could be largely attributed to inadequate reporting of detailed methods, no or limited verification by authors, and the production and use of unvalidated antibodies. The goal of this guideline article is to review best practices concerning commonly used techniques involving antibodies, including immunoblotting, immunohistochemistry, and flow cytometry. Awareness and integration of best practices will increase the rigor and reproducibility of these techniques and elevate the quality of physiology research.

Keywords: Western blotting; flow cytometry; immunoblotting; immunofluorescence; immunohistochemistry.

MeSH terms

  • Antibodies*
  • Antibody Specificity
  • Flow Cytometry
  • Immunohistochemistry
  • Reproducibility of Results

Substances

  • Antibodies