ArticleHistochemistry and cell biology2016
A microwave antigen retrieval method using two heating steps for enhanced immunostaining on aldehyde-fixed paraffin-embedded tissue sections.
Article in Histochemistry and cell biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- A new efficient immunoprotocol to detect chromosomal/nuclear proteins along with repetitive DNA in squash preparations of formalin-fixed, long-stored root tips.Plant methods · 2025Article
- Guidelines on antibody use in physiology research.American journal of physiology. Renal physiology · 2024Article
- Article
- Generation of iPSC-derived human forebrain organoids assembling bilateral eye primordia.Nature protocols · 2023Review
- Redeployment of odontode gene regulatory network underlies dermal denticle formation and evolution in suckermouth armored catfish.Scientific reports · 2022Article
- Oyster Heat Shock Protein 70 Plays a Role in Binding of Human Noroviruses.Applied and environmental microbiology · 2021Article
- Multicolor Immunofluorescence Staining of Paraffin-Embedded Human Bone Marrow Sections.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Endocycle-related tubular cell hypertrophy and progenitor proliferation recover renal function after acute kidney injury.Nature communications · 2018Article
- Mechanical loading modulates heterotopic ossification in calcific tendinopathy through the mTORC1 signaling pathway.Molecular medicine reports · 2017Article
- Immunohistochemical techniques for the human inner ear.Histochemistry and cell biology · 2016Review
- Microwave-Assisted Tissue Preparation for Rapid Fixation, Decalcification, Antigen Retrieval, Cryosectioning, and Immunostaining.International journal of cell biology · 2016Review
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Authors and funding
5 authors.
Funding
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Abstract
Antigen retrieval is an immunohistochemical procedure that results in better exposure of target antigens in aldehyde-fixed, paraffin-embedded tissue sections to antibodies. However, the commercially recommended or conventional protocols for antigen retrieval do not always succeed in expressing the target antigen. Here, an improved method was developed for antigen retrieval from aldehyde-fixed, paraffin-embedded histological sections. Proliferating cell nuclear antigen (PCNA), tight junction proteins Claudin-2 and Claudin-7, and water channel aquaporins in kidney tissue were selected as test antigens. Typically, PCNA and Claudin-2 and Claudin-7 show negative, weak, or nonspecific immunoreactions with conventional antigen retrieval methods using microwave heating. In the present study, microwave heating was performed twice with an interval of 30 min between the two steps to allow the buffer solution to cool. Sodium citrate buffer (10 mM sodium citrate, pH 6.0) was used for PCNA, and Tris-EDTA buffer (10 mM Tris, 1 mM EDTA, pH 9.0) was used for the Claudins. Compared with conventionally prepared tissues, the tissues exhibited both enhanced and specific immunostaining, and well-preserved morphology. In conclusion, the conventional protocol could be supplemented with a second microwave heating step to improve the expression of antigens that do not respond well to the conventional method.
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