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Immunofluorescent Staining of Adult Murine Paraffin-Embedded Skeletal Tissue

Neta Felsenthal, Elazar Zelzer

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Immunohistochemistry, or immunolabeling, is a key method for the identification of protein expression and localization. Successful detection relies on a low signal-to-noise ratio, which is affected greatly by antibody specificity as well as the staining protocol. Immunohistochemistry in the mouse is challenging, particularly in adult skeletal tissue, due to the need for long decalcification, high autofluorescence and high levels of endogenous peroxidase. Here, we describe a highly sensitive protocol for protein detection in decalcified paraffin-embedded sections from adult mouse skeletal tissue. By using four levels of amplification, this method allows for the identification of even low-abundance proteins.

Original languageEnglish
Title of host publicationSkeletal Development and Repair
Subtitle of host publicationMethods and Protocols
EditorsMatthew J. Hilton
PublisherHumana Press
Chapter19
Pages337-344
Number of pages8
ISBN (Electronic)978-1-0716-1028-2
ISBN (Print)978-1-0716-1027-5
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2230
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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