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Histochemical and Fluorochrome-Based Detection of β-Galactosidase

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Recombinant Protein Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 63))

Abstract

Bacterial β-galactosidase (β-gal the LacZ gene product) has been used extensively as a reporter gene in a wide variety of systems. It has been especially useful in transgenic and chimeric mouse studies. In such whole-animal applications, the experimental goals often include the evaluation of the spatial and tissue specific patterns of reporter gene expression in situ. In such instances, the LacZ reporter gene has the advantage over other reporters in that in situ histochemical staining for β-gal activity is relatively simple and sensitive. Additionally, fluorogenic substrates for β-gal are available that can be used to identify and isolate living cells expressing the β-gal reporter. Here, we discuss two protocols for localizing β-gal activity, one for staining fixed specimens, and the other for staining having cells. Each of these techniques has unique advantages and limitations, but together they offer a wide range of possibilities for taking advantage of β-gal as a reporter enzyme in a variety of applications.

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Author information

Authors and Affiliations

  1. Department of Biology, University of Pennsylvania, Philadelphia, PA

    Ruth Sullivan & Cecilia W. Lo

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  1. Ruth Sullivan
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  2. Cecilia W. Lo
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Editors and Affiliations

  1. Thomas Jefferson University, Philadelphia, PA

    Rocky S. Tuan

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© 1997 Humana Press Inc., Totowa, NJ

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Sullivan, R., Lo, C.W. (1997). Histochemical and Fluorochrome-Based Detection of β-Galactosidase. In: Tuan, R.S. (eds) Recombinant Protein Protocols. Methods in Molecular Biology™, vol 63. Humana Press. https://doi.org/10.1385/0-89603-481-X:229

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  • DOI: https://doi.org/10.1385/0-89603-481-X:229

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-481-5

  • Online ISBN: 978-1-59259-549-5

  • eBook Packages: Springer Protocols

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