Antibody validation of immunohistochemistry for biomarker discovery: recommendations of a consortium of academic and pharmaceutical based histopathology researchers

Methods. 2014 Nov;70(1):34-8. doi: 10.1016/j.ymeth.2014.01.018. Epub 2014 Feb 11.


As biomarker discovery takes centre-stage, the role of immunohistochemistry within that process is increasing. At the same time, the number of antibodies being produced for "research use" continues to rise and it is important that antibodies to be used as biomarkers are validated for specificity and sensitivity before use. This guideline seeks to provide a stepwise approach for the validation of an antibody for immunohistochemical assays, reflecting the views of a consortium of academic and pharmaceutical based histopathology researchers. We propose that antibodies are placed into a tier system, level 1-3, based on evidence of their usage in immunohistochemistry, and that the degree of validation required is proportionate to their place on that tier.

Keywords: Antibody validation; Biomarker; Guidelines; Histopathology; Immunohistochemistry.

Publication types

  • Guideline
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / chemistry*
  • Biomarkers / chemistry
  • Biomarkers / metabolism*
  • Biomarkers, Tumor / metabolism
  • Biomedical Research / methods
  • Cell Line
  • Chemistry, Pharmaceutical / methods
  • Epitopes / chemistry
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Immunohistochemistry / methods*
  • Neoplasms / immunology
  • Neoplasms / metabolism
  • Phenotype
  • Proteins / chemistry*
  • RNA, Messenger / metabolism
  • Reproducibility of Results


  • Antibodies
  • Biomarkers
  • Biomarkers, Tumor
  • Epitopes
  • Proteins
  • RNA, Messenger