A new simplified catalyzed signal amplification system for minimizing non-specific staining in tissues with supersensitive immunohistochemistry

DOI Web Site PubMed 被引用文献4件 参考文献73件
  • Hasui Kazuhisa
    Department of Structural Cell Biology, Field of Musculo-Skeletal Disorder, Course of Advanced Therapeutics, Kagoshima University Graduate School of Medical and Dental Sciences
  • Murata Fusayoshi
    Department of Structural Cell Biology, Field of Musculo-Skeletal Disorder, Course of Advanced Therapeutics, Kagoshima University Graduate School of Medical and Dental Sciences

この論文をさがす

抄録

We investigated non-specific staining in a catalyzed reporter deposition (CARD) reaction and improved its blocking methods in supersensitive immunohistochemistry, based on our simplified catalyzed signal amplification (CSA) system (Hasui et al. 2002). In the CARD reaction using biotinyl tyramide, non-specific staining could be reduced by pretreatment with a casein solution or 3% bovine serum albumin (BSA)-phosphate buffer saline (PBS) with 0.1% Tween 20. In the CARD reaction using FITC-labeled tyramide, non-specific staining could be blocked by pretreatment with 0.3% BSA-PBS with 0.1% Tween 20 or 3% polyethylene glycol-PBS with 01% Tween 20. Thus, our new simplified CSA system features: 1) destruction of the endogenous peroxidase activity; 2) blocking of the nonspecific reaction of the primary antibody; 3) a primary antibody reaction; 4) blocking of the non-specific reaction of the polymer reagent by casein treatment; 5) a polymer reaction; 6) blocking of the non-specific reaction of CARD reaction by casein treatment; 7) a CARD reaction; and 8) detection of deposited tyramide. This new system proved useful for detecting an extremely low amount of antigen in the endogenous biotin-rich tissues such as the gastrointestinal tract and liver. By this method, the Ki67 antigen in the G1 phase cell cycle could be detected and a metabolic disorder of the Ki67 antigen was implicated in a carcinoid tumor in the stomach. We believe that this new simplified CSA system represents a new standard of supersensitive immunohistochemistry for use in light-microscopic investigation.

収録刊行物

被引用文献 (4)*注記

もっと見る

参考文献 (73)*注記

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ