title: Proper application of antibodies for immunohistochemical detection: antibody crimes and how to prevent them.
abstract: For several decades antibodies raised against specific proteins, peptides, or peptide epitopes have proven to be versatile and very powerful tools to demonstrate molecular identity in cells and tissues. New techniques of immunohistochemistry and immunofluorescence have improved both the optical resolution of such protein identification as well as its sensitivity, particularly through the use of amplification methodology. However, this improved sensitivity has also increased the risks of false-positive and false-negative staining and thereby raised the necessity for proper and adequate controls. In this review, the authors draw on many years of experience to illuminate many of the more common errors and problematic issues in immunohistochemistry, and how these may be avoided. A key factor in all of this is that techniques need to be properly documented and especially antibodies and procedures must be adequately described. Antibodies are a valuable and shared resource within the scientific community; it is essential therefore that mistakes involving antibodies and their controls are not perpetuated through inadequate reporting in the literature.
authors.affiliation: Leibniz Institute for Farm Animal Biology (R.I.), 18196 Dummerstorf, Germany; School of Molecular and Biomedical Science (R.I.), University of Adelaide, SA5005, Australia; Department of Animal Sciences (K.T.), Wageningen University, 6709 WD Wageningen, The Netherlands; and Department of Biology (G.E.H.), Morgan State University, Baltimore, Maryland 21251.
authors.forename: Richard
authors.initials: R
authors.lastname: Ivell
authors.forename: Katja
authors.initials: K
authors.lastname: Teerds
authors.forename: Gloria E
authors.initials: GE
authors.lastname: Hoffman
mesh_terms: D000327:Adsorption; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D000941:Antigens / Q000737:chemistry; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry / Q000295:instrumentation* / Q000379:methods*; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence / Q000379:methods; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000906:Antibodies; D000941:Antigens; D000939:Epitopes