An Improved Protocol of Biotinylated Tyramine-based Immunohistochemistry Minimizing Nonspecific Background Staining
- Kim, Seok Hyung; Shin, Young Kee; Lee, Kyung Mee; Lee, Jung Sun; Yun, Ji Hye; Lee, Sun Myung
- Issue Date
- Histochemical Society
- J Histochem Cytochem; Vol.51, No.1; pp.129–132
- An immunohistochemical method using biotinyl tyramine was recently introduced
to amplify weak staining signals. Despite its high sensitivity, however, tyraminebased
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.
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