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Journal of Histochemistry and Cytochemistry, Vol. 51, 129-132, January 2003, Copyright © 2003, The Histochemical Society, Inc.

BRIEF REPORT

An Improved Protocol of Biotinylated Tyramine-based Immunohistochemistry Minimizing Nonspecific Background Staining

Seok Hyung Kima,b, Young Kee Shina,b, Kyung Mee Leeb, Jung Sun Leeb, Ji Hye Yunb, and Sun Myung Leeb
a Department of Pathology, Seoul National University College of Medicine, Seoul, Korea
b DiNonA, Inc., Seoul, Korea

Correspondence to: Seok Hyung Kim, DiNonA Inc., 65 Umyeon-dong, Seo-cho Gu, Seoul, South Korea 137-140. E-mail: platoshkim@freechal.com

An immunohistochemical method using biotinyl tyramine was recently introduced to amplify weak staining signals. Despite its high sensitivity, however, tyramine-based immunostaining has been limited by its increased background staining. In this study, to develop an improved protocol of biotinyl tyramine-based immunohistochemistry minimizing the background staining, we determined which staining steps lead to the nonspecific reaction and the most appropriate blocking agents for background-provoking steps. Trypton casein peptone and distilled water with Tween-20 were shown to be most effective as a blocking agent and a rinsing solution, respectively. In conclusion, we developed an optimized protocol for biotinyl tyramine-based immunohistochemistry with minimal background staining. (J Histochem Cytochem 51:129–132, 2003)

Key Words: immunohistochemistry, tyramine, background staining


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