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Immunostaining of Skeletal Tissues

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Osteoporosis and Osteoarthritis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2221))

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Abstract

Immunostaining is the process of identifying proteins in tissue sections by incubating the sample with antibodies specific to the protein of interest, then visualizing the bound antibody using a chromogen (immunohistochemistry or IHC) or fluorescence (immunofluorescence or IF). Unlike in situ hybridization, which identifies gene transcripts in cells, immunostaining identifies the products themselves and provides information about their localization within cells (nuclear, cytoplasmic, or membrane) or extracellular matrix. This can be particularly important in the context of bone and cartilage because they contain many cell types as well as matrix components, each with distinct protein expression patterns. As the number of antibodies continues to grow, this technique has become vital for research laboratories studying the skeleton. Here, we describe a detailed protocol for antibody-based in situ analysis of bone and associated tissues, addressing specific issues associated with staining of hard and matrix-rich tissues.

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Author information

Author notes

  1. Crystal Idleburg and Madelyn R. Lorenz contributed equally to this work.

Authors and Affiliations

  1. Musculoskeletal Research Center, Histology and Morphometry Core, Washington University, St. Louis, MO, USA

    Crystal Idleburg, Madelyn R. Lorenz, Elizabeth N. DeLassus, Erica L. Scheller & Deborah J. Veis

  2. Department of Orthopedics, Washington University, St. Louis, MO, USA

    Crystal Idleburg & Elizabeth N. DeLassus

  3. Department of Medicine, Division of Bone and Mineral Diseases, Washington University, St. Louis, MO, USA

    Madelyn R. Lorenz, Erica L. Scheller & Deborah J. Veis

  4. Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA

    Deborah J. Veis

Authors
  1. Crystal Idleburg
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  2. Madelyn R. Lorenz
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  3. Elizabeth N. DeLassus
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  4. Erica L. Scheller
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  5. Deborah J. Veis
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Corresponding author

Correspondence to Deborah J. Veis .

Editor information

Editors and Affiliations

  1. Stem Cell Therapy and Skeletal Regeneration Lab, Mayo Clinic, Rochester, MN, USA

    Andre J. van Wijnen

  2. Stem Cell Therapy and Skeletal Regeneration Lab, Mayo Clinic, Rochester, MN, USA

    Marina S. Ganshina

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Idleburg, C., Lorenz, M.R., DeLassus, E.N., Scheller, E.L., Veis, D.J. (2021). Immunostaining of Skeletal Tissues. In: van Wijnen, A.J., Ganshina, M.S. (eds) Osteoporosis and Osteoarthritis. Methods in Molecular Biology, vol 2221. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0989-7_15

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  • DOI: https://doi.org/10.1007/978-1-0716-0989-7_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0988-0

  • Online ISBN: 978-1-0716-0989-7

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