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Immunohistochemistry and In Situ Hybridization in the Developing Chicken Brain

  • Richard P. TuckerEmail author
  • Tatsuto Ishimaru
  • Qizhi Gong
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2047)

Abstract

One of the first steps in studies of gene function is the spatiotemporal analysis of patterns of gene expression. Indirect immunohistochemistry is a method that allows the detection of a protein of interest by incubating a histological section with an antibody or antiserum raised against the protein, and then localizing this primary antibody with a tagged secondary antibody. To determine the cellular source of a protein of interest, or if a specific antibody is not available, specific transcripts can be localized using in situ hybridization. A histological section is incubated with a labeled RNA probe that is complementary to the target transcript; after hybridization with the target transcript the labeled RNA probe can be identified with an antibody. Here we describe materials and methods used to perform basic indirect immunohistochemistry and in situ hybridization on frozen sections through the developing chicken brain, emphasizing controls and potential problems that may be encountered.

Keywords

Immunohistochemistry In situ hybridization Riboprobe Antibody Fluorescence Protocol Technique Cryosection 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Richard P. Tucker
    • 1
    Email author
  • Tatsuto Ishimaru
    • 1
  • Qizhi Gong
    • 1
  1. 1.Department of Cell Biology and Human AnatomyUniversity of California, DavisDavisUSA

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