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Immunohistochemical detection of beta-galactosidase or green fluorescent protein on tissue sections

Item Type:Article
Title:Immunohistochemical detection of beta-galactosidase or green fluorescent protein on tissue sections
Creators Name:Seymour, P.A. and Sander, M.
Abstract:With the recent advances in mouse genetics, it is now possible to mark specific cell types genetically in vivo and to study the fate of cells during development and adulthood. Cells are labeled and followed in vivo through the stable expression of reporter genes in particular cell types. The two most commonly used reporter genes are LacZ, which encodes the enzyme beta-galactosidase (beta-gal), and green fluorescent protein (GFP). beta-Gal expression can be detected enzymatically, using 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) as a substrate, and GFP can be directly visualized by fluorescence microscopy. However, with single detection of beta-gal or GFP, it is often impossible to determine whether expression of the reporter protein is restricted to a particular cell type. To ascertain the identity of individual cells within a multicellular tissue, beta-gal or GFP proteins must be visualized in conjunction with additional cellular markers. For such experiments, specific antibodies raised against beta-gal or GFP can be used in coimmunofluorescence analyses. Such double-staining analyses on tissue sections are a powerful tool to study transgene expression or to trace cells in multicellular tissues.
Keywords:Reporter Gene, Mouse, Green Fluorescent Protein, LacZ, ß-Galactosidase, Immunofluorescence, Immunohistochemistry, Fluorochrome, Antibody, Animals, Mice
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:Reporter genes: a practical guide
Publisher:Springer / Humana Press
Page Range:13-23
Number of Pages:158
Date:13 September 2007
Official Publication:https://doi.org/10.1007/978-1-59745-549-7_2
PubMed:View item in PubMed

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