An improved protocol of biotinylated tyramine-based immunohistochemistry minimizing nonspecific background staining. |
| |
Authors: | Seok Hyung Kim Young Kee Shin Kyung Mee Lee Jung Sun Lee Ji Hye Yun Sun Myung Lee |
| |
Affiliation: | Department of Pathology, Seoul National University College of Medicine, Seoul, Korea. platoshkim@freechal.com |
| |
Abstract: | 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. |
| |
Keywords: | |
|
|