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論文
- タイトル
- タイトル（英）
- Dual enhancement of triple immunofluorescence using two antibodies from the same species
- 参照URL
- https://researchmap.jp/brain-pathology/published_papers/1663056
- 著者
- 著者（英）
- A Nakamura,T Uchihara
- 担当区分
- 概要
- 概要（英）
- Triple immunofluorescence method with two mouse monoclonal antibodies and another rabbit polyclonal antibody was established with catalyzed reporter deposition (CARD) amplification on thick floating sections from the rat cerebellum. One of the monoclonal antibodies (anti-calbindin). diluted maximally. probed with anti-mouse IgG-horseradish peroxidase (HRP) and amplified with Cy5-conjugated tyramide, immunolabeled cerebellar Purkinje cells and their arborization. Subsequently, a rabbit polyclonal IgG (anti-glial fibrillary acidic protein (anti-GFAP)), probed with anti-rabbit IgG-HRP, amplified with biotin-tyramide and visualized with fluorescein-isothiocyanate (FITC)-streptavidin. immunolabeled Bergmann's glia. Another mouse monoclonal IgG (anti-SNAP25), probed with anti-mouse IgG-rhodamine without CARD amplification, selectively visualized synaptic sites, because the maximal dilution of the other monoclonal antibody (anti-calbindin) was below the detection threshold of this anti-mouse IgG-rhodamine. Separation of the two signals (calbindin and SNAP25), each detected through mouse monoclonal antibody, was then based on the difference of sensitivity either with or without CARD amplification. Triple immunofluorescence is possible when just one of the three primary antibodies is from different species. Intensification of two of the three signals provides further advantages to examine immunolocalization of multiple epitopes on histological sections. (C) 2004 Elsevier B.V. All rights reserved.
- 出版者・発行元
- 出版者・発行元（英）
- ELSEVIER SCIENCE BV
- 誌名
- 誌名（英）
- JOURNAL OF NEUROSCIENCE METHODS
- 巻
- 135
- 号
- 1-2
- 開始ページ
- 67
- 終了ページ
- 70
- 出版年月
- 2004年5月
- 査読の有無
- 査読有り
- 招待の有無
- 掲載種別
- 研究論文（学術雑誌）
- ISSN
- 0165-0270
- DOI URL
- https://doi.org/10.1016/j.jneumeth.2003.12.003
- 共同研究・競争的資金等の研究課題
研究者
内原 俊記
(ウチハラ トシキ)