Immunohistochemical Detection of β-Galactosidase or Green Fluorescent Protein on Tissue Sections
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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 β-galactosidase (β-gal), and green fluorescent protein (GFP). β-Gal expression can be detected enzymatically, using 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal) as a substrate, and GFP can be directly visualized by fluorescence microscopy. However, with single detection of β-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, β-gal or GFP proteins must be visualized in conjunction with additional cellular markers. For such experiments, specific antibodies raised against β-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.
- Branda, C. S. and Dymecki, S. M. (2004) Talking about a revolution: the impact of site-specific recombinases on genetic analyses in mice. Dev. Cell 6, 7–28. CrossRef
- Soriano, P. (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat. Genet. 21, 70–71. CrossRef
- Lobe, C. G., Koop, K. E., Kreppner, W., Lomeli, H., Gertsenstein, M., and Nagy, A. (1999) Z/AP, a double reporter for cre-mediated recombination. Dev. Biol. 208, 281–292. CrossRef
- Novak, A., Guo, C., Yang, W., Nagy, A., and Lobe, C. G. (2000) Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cremediated excision. Genesis 28, 147–155. CrossRef
- Mao, X., Fujiwara, Y., Chapdelaine, A., Yang, H., and Orkin, S. H. (2001) Activation of EGFP expression by Cre-mediated excision in a new ROSA26 reporter mouse strain. Blood 97, 324–326. CrossRef
- Hadjantonakis, A. K., Dickinson, M. E., Fraser, S. E., and Papaioannou, V. E. (2003) Technicolour transgenics: imaging tools for functional genomics in the mouse. Nat. Rev. Genet. 4, 613–625. CrossRef
- Mombaerts, P., Wang, F., Dulac, C., et al. (1996) Visualizing an olfactory sensory map. Cell 87, 675–686. CrossRef
- Giepmans, B. N., Adams, S. R., Ellisman, M. H., and Tsien, R. Y. (2006) The fluorescent toolbox for assessing protein location and function. Science 312, 217–224. CrossRef
- Ausubel, F., Brent, R., Kingston, R. E., et al., eds. (1987) Current Protocols in Molecular Biology, vol. 3, John Wiley & Sons, New York, 14.1.1–14.6.8.
- Brown, R. W. and Chirala, R. (1995) Utility of microwave-citrate antigen retrieval in diagnostic immunohistochemistry. Mod. Pathol. 8, 515–520.
- Shi, S. R., Cote, R. J., and Taylor, C. R. (1997) Antigen retrieval immunohistochemistry: past, present, and future. J. Histochem. Cytochem. 45, 327–343.
- Shi, S. R., Chaiwun, B., Young, L., Cote, R. J., and Taylor, C. R. (1993) Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections. J. Histochem. Cytochem. 41, 1599–1604.
- Immunohistochemical Detection of β-Galactosidase or Green Fluorescent Protein on Tissue Sections
- Book Title
- Reporter Genes
- Book Subtitle
- A Practical Guide
- pp 13-23
- Print ISBN
- Online ISBN
- Series Title
- Methods in Molecular Biology
- Series Volume
- Series ISSN
- Humana Press
- Copyright Holder
- Humana Press Inc.
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- Reporter gene
- green fluorescent protein
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