An improved protocol of biotinylated tyramine-based immunohistochemistry minimizing nonspecific background staining.
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.
Kim
Seok Hyung
SH
Department of Pathology, Seoul National University College of Medicine, Seoul, Korea. platoshkim@freechal.com
Shin
Young Kee
YK
Lee
Kyung Mee
KM
Lee
Jung Sun
JS
Yun
Ji Hye
JH
Lee
Sun Myung
SM
D001710:Biotin / Q000031:analogs & derivatives*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008198:Lymph Nodes / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D014439:Tyramine / Q000031:analogs & derivatives*
C111551:biotinyltyramide; D001710:Biotin; D014439:Tyramine