Amplification of Immunostaining Intensity by the Peroxidase-Antiperoxidase, Avidin-Biotin-Peroxidase Complex or Catalyzed Signal Amplification Method Gives Erroneous Antigen Content in Sections
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概要
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The peroxidase-antiperoxidase (PAP), avidinbiotin-peroxidase complex (ABC) and catalyzed signal amplification (CSA) methods have been used for quantitative or semiquantitative immunohistochemical analysis. It is however unclear whether intensity of immunostaning in sections amplified by using the PAP, ABC or CSA method reflects an antigen content. Thus, relationship between the intensity and the content was examined in antigen-immobilized nitrocellulose filters or serial sections stained by using the PAP, ABC or CSA method. In the present study, we selected cytochromes P-450 2B1/2 (CYPB1/2), because the immunostaining intensity of the enzymes detected with the indirect immunoperoxidase (IPO) method under saturation conditions is shown to be proportional to the antigen content in sections. For the IPO method, the intensity in filters or sections increased with increasing concentration of anti-CYP2B1/2 antibody, and then saturated. However, a bell-shaped relationship was observed between the intensity and the primary antibody concentration in filters or sections stained by using the PAP, ABC or CSA method. Furthermore, the amplified intensity in filters or sections was disproportionate to the antigen content. Thus, erroneous CYP2B1/2 content was obtained from sections in which the intensity was amplified by the PAP, ABC or CSA method. It is therefore unlikely that the PAP, ABC and CSA methods are suitable for the measurement of antigen content in sections by quantitative immunohistochemistry.
- 日本組織細胞化学会の論文
著者
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WATANABE Jun
Department of Cardiovascular Medicine, Tohoku University Graduate School of Medicine
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Watanabe J
Department Of Anatomy & Cell Science Kansai Medical University
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TAKAMORI Yasuharu
Department of Anatomy and Cell Science, Kansai Medical University
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Mondo Hiroko
Department of Anatomy and Cell Science, Kansai Medical University
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Tamura Yasuhisa
Department Of Anatomy And Cell Science Kansai Medical University
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TAKEDA Kazuo
Department of Medicine, Kyoto Prefectural University of Medicine
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Watanabe Junichi
Aska Pharmaceutical Co. Ltd. Research & Development Division
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Takeda Koichi
Department Of Pathology Wakayama Medical University
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Kanamura Shinsuke
Department of Anatomy, Kansai Medical University
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Kanamura Shinsuke
Department Of Anatomy Kansai Medical University
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Mondo Hiroko
Department Of Anatomy And Cell Science Kansai Medical University
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Watanabe Jun
Department Of Pathology School Of Medicine Kitasato University
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Watanabe Jun
Department Of Anatomy & Cell Science Kansai Medical University
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Tabuse Katsuyoshi
Department Of Surgery National Hospital Organization Osaka Minami Medical Center
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Kanamura S
Kansai Medical Univ.
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Takeda Kazuo
Department Of Biology School Of Medicine University Of Occupational And Environmental Health
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Takamori Yasuharu
Department Of Anatomy Kansai Medical University
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Takamori Yasuharu
Department Of Anatomy And Cell Science Kansai Medical University
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