Antibodies for immunolabeling by light and electron microscopy: not for the faint hearted

Gareth Griffiths*, John Milton Lucocq

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

39 Citations (Scopus)
1 Downloads (Pure)

Abstract

Reliable antibodies represent crucial tools in the arsenal of the cell biologist and using them to localize antigens for immunocytochemistry is one of their most important applications. However, antibody-antigen interactions are much more complex and unpredictable than suggested by the old 'lock and key' analogy, and the goal of trying to prove that an antibody is specific is far more difficult than is generally appreciated. Here, we discuss the problems associated with the very complicated issue of trying to establish that an antibody (and the results obtained with it) is specific for the immunolabeling approaches used in light or electron microscopy. We discuss the increasing awareness that significant numbers of commercial antibodies are often not up to the quality required. We provide guidelines for choosing and testing antibodies in immuno-EM. Finally, we describe how quantitative EM methods can be used to identify reproducible patterns of antibody labeling and also extract specific labeling distributions.

Original languageEnglish
Pages (from-to)347-360
Number of pages14
JournalHistochemistry and Cell Biology
Volume142
Issue number4
Early online date24 Aug 2014
DOIs
Publication statusPublished - Oct 2014

Keywords

  • Antibodies/Specificity
  • Immunocytochemistry
  • Stereology/Quantitation
  • Commercial antibodies
  • Light microscopy/EM
  • Immunoelectron microscopy
  • Cross-reactivity
  • Specificity
  • Protein
  • Immunohistochemistry
  • Receptors
  • Channel
  • Complex

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