Antibody Specificity Profiling Using Protein Microarrays

Methods Mol Biol. 2018;1785:223-229. doi: 10.1007/978-1-4939-7841-0_14.


Antibodies are the most widely used reagent for isolation and detection of specific proteins. However, using antibodies that are not highly specific in these studies can generate inaccurate and misleading data. Protein microarrays offer a platform by which antibody cross-reactivity against a broad range of cellular antigens can be simultaneously and quantitatively profiled. This protocol describes in detail the process of array pretreatment, antibody binding, washing, scanning and quantitative analysis of antibody specificity.

Keywords: Antibody specificity; Cross-reactivity; Monoclonal antibodies; Protein microarray.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies / genetics
  • Antibodies / immunology
  • Antibody Specificity / immunology*
  • Antigens / genetics
  • Antigens / immunology
  • Humans
  • Protein Array Analysis / methods*
  • Proteomics / methods*


  • Antibodies
  • Antigens