title: The enhanced reactivity of endogenous biotin-like molecules by antigen retrieval procedures and signal amplification with tyramine.
abstract: In diagnostic pathology and immunocytochemical research, immunohistochemical techniques using the streptavidin-biotin-peroxidase system have played an extremely valuable role. This system, based on the high affinity of streptavidin for biotin, may, however, provoke false positive results because of endogenous streptavidin-binding sites in human tissues. With the advent of the antigen retrieval procedure and signal amplification method, this problem can be serious enough to cause mistakes in interpreting immunohistochemical staining results. Therefore, we examined the distribution of endogenous biotin-like molecules in various human tissues and the influence of various antigen retrieval procedures with or without signal amplification using biotinylated tyramine to reveal these biotin-like activities. We observed that endogenous biotin-like molecules were present in a wide range of tissues, and their activity was markedly enhanced by employing antigen retrieval procedures or signal amplification. Furthermore, the extent to which the activity of endogenous biotin-like activities was enhanced depended on the kinds of antigen retrieval procedures and signal amplification employed. Pressure cooking and tyramine amplification with microwave heating showed the highest activities. These results show that the antigen retrieval procedures and signal amplification with tyramine can enhance the activity of endogenous biotin or biotin-like molecules as well as antigenicity, which can be a pitfall in the interpretation of immunohistochemical data.
authors.affiliation: Department of Pathology, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, Seoul 110-799, South Korea.
authors.forename: Seok Hyung
authors.initials: SH
authors.lastname: Kim
authors.forename: Kyeong Cheon
authors.initials: KC
authors.lastname: Jung
authors.forename: Young Kee
authors.initials: YK
authors.lastname: Shin
authors.forename: Kyung Mee
authors.initials: KM
authors.lastname: Lee
authors.forename: Young S
authors.initials: YS
authors.lastname: Park
authors.forename: Yoon La
authors.initials: YL
authors.lastname: Choi
authors.forename: Kwon Ik
authors.initials: KI
authors.lastname: Oh
authors.forename: Min Kyung
authors.initials: MK
authors.lastname: Kim
authors.forename: Doo Hyun
authors.initials: DH
authors.lastname: Chung
authors.forename: Hyung Geun
authors.initials: HG
authors.lastname: Son
authors.forename: Seong Hoe
authors.initials: SH
authors.lastname: Park
mesh_terms: D000818:Animals; D000941:Antigens / Q000737:chemistry*; D001360:Avidin / Q000737:chemistry; D001710:Biotin / Q000037:antagonists & inhibitors / Q000502:physiology*; D004396:Coloring Agents; D006735:Horseradish Peroxidase; D006358:Hot Temperature; D006801:Humans; D006868:Hydrolysis; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D008872:Microwaves; D011312:Pressure; D014357:Trypsin; D014439:Tyramine / Q000737:chemistry*
chemical_list: D000941:Antigens; D004396:Coloring Agents; D001360:Avidin; D001710:Biotin; D006735:Horseradish Peroxidase; D014357:Trypsin; D014439:Tyramine