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Hypothesis for the mechanism for heat-induced antigen retrieval occurring on fresh frozen sections without formalin-fixation in immunohistochemistry

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Abstract

The mechanism involved in heat-induced antigen retrieval (AR) remains unproven but probably utilizes the breaking of formalin-induced cross-linkages. We investigated the effectiveness of heat-induced AR on immunohistochemistry and dot-blot analysis using rat uterus tissue sections and protein extracts without formalin-fixation. The unfixed frozen sections, which did not show immunostaining with nine antibodies, were clearly stained after heating the sections. In the dot-blot analysis, the immunoblot sensitivity of detection was greatly enhanced by heating the protein-blotted membrane. These results indicate that other mechanisms of breaking formalin-induced cross-linkages may be present. We propose that one of the other mechanisms for heat-induced AR is that accessibility to the target epitopes of antigenic proteins is limited by natural steric barriers even in the fresh state caused by the antigenic protein itself.

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Acknowledgments

We thank Dr. Shoichiro Sugai, Dr. Yoshifumi Miyakawa, Dr. Haruko Koizumi and Dr. Hisashi Iwatsuka for their support and advice.

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Authors and Affiliations

  1. Department of Pathology, Tokai University School of Medicine, 143-Shimokasuya, Isehara, Kanagawa, 259-1193, Japan

    Kochi Kakimoto, Susumu Takekoshi, Katsuhiro Miyajima & R. Yoshiyuki Osamura

  2. Toxicology Research Laboratories, Central Pharmaceutical Research Institute, Japan Tobacco Inc., 23-Naganuki, Hadano, Kanagawa, 257-0024, Japan

    Kochi Kakimoto & Katsuhiro Miyajima

Authors
  1. Kochi Kakimoto
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  2. Susumu Takekoshi
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  3. Katsuhiro Miyajima
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  4. R. Yoshiyuki Osamura
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Correspondence to Susumu Takekoshi.

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Kakimoto, K., Takekoshi, S., Miyajima, K. et al. Hypothesis for the mechanism for heat-induced antigen retrieval occurring on fresh frozen sections without formalin-fixation in immunohistochemistry. J Mol Hist 39, 389–399 (2008). https://doi.org/10.1007/s10735-008-9177-y

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  • DOI: https://doi.org/10.1007/s10735-008-9177-y

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