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. 2025:2885:461-477.
doi: 10.1007/978-1-0716-4306-8_24.

Immunostaining of Skeletal Tissues

Susan M Clark  1 , Cosimo De Bari  1   2 , Anke J Roelofs  3   4
Affiliations

Immunostaining of Skeletal Tissues

Susan M Clark et al. Methods Mol Biol. 2025.

Abstract

Immunohistochemistry (IHC) is a routinely used technique in clinical diagnosis of pathological conditions and in basic and translational research. It combines anatomical, immunological, and biochemical methods and relies on the specific binding of an antibody to an antigen. Using the technique with mineralized tissues is more challenging than with soft tissues. Demineralizing the samples allows embedding in paraffin wax, and also facilitates cryosectioning. This chapter describes methods for IHC on formaldehyde-fixed, demineralized, paraffin-embedded, or frozen sections to detect antigens in skeletal tissues. We include a protocol using a streptavidin-based amplification method for immunofluorescence staining.

Keywords: Antigen retrieval; Cryosection; Demineralization; Immunofluorescence; Immunohistochemistry; Paraffin section; Skeletal tissues.

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