The enhanced reactivity of endogenous biotin-like molecules by antigen retrieval procedures and signal amplification with tyramine.
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.
Kim
Seok Hyung
SH
Department of Pathology, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, Seoul 110-799, South Korea.
Jung
Kyeong Cheon
KC
Shin
Young Kee
YK
Lee
Kyung Mee
KM
Park
Young S
YS
Choi
Yoon La
YL
Oh
Kwon Ik
KI
Kim
Min Kyung
MK
Chung
Doo Hyun
DH
Son
Hyung Geun
HG
Park
Seong Hoe
SH
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*
D000941:Antigens; D004396:Coloring Agents; D001360:Avidin; D001710:Biotin; D006735:Horseradish Peroxidase; D014357:Trypsin; D014439:Tyramine