Immunohistochemistry and In Situ Hybridization in the Developing Chicken Brain

Richard P. Tucker, Tatsuto Ishimaru, Qizhi Gong

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages421-437
Number of pages17
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2047
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Antibody
  • Cryosection
  • Fluorescence
  • Immunohistochemistry
  • In situ hybridization
  • Protocol
  • Riboprobe
  • Technique

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Tucker, R. P., Ishimaru, T., & Gong, Q. (2020). Immunohistochemistry and In Situ Hybridization in the Developing Chicken Brain. In Methods in Molecular Biology (pp. 421-437). (Methods in Molecular Biology; Vol. 2047). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9732-9_24