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Methods for Double Detection of Gene Expression

Combined In Situ Hybridization and Immunocytochemistry or Histochemistry

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Recombinant Protein Protocols

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

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Abstract

The distribution of two different molecules can be analyzed within the same embryo using the procedures described below. The protocol for combined whole mount in situ hybridization and immunocytochemistry allows for simultaneous detection of mRNA and protein. The protocol for combined whole mount in situ hybridization and β-galactosidase staining allows for simultaneous detection of mRNA and transgene-directed β-galactosidase expression. Simultaneous detection allows for the most direct comparison of expression patterns. These procedures are derived from protocols used in Drosophila (1) and mice (2,3).

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References

  1. Cubas, P., de Celis, J.-F., Campuzano, S., and Modolell, J. (1991) Proneural clusters of achaete-scute expression and the generation of sensory organs in the Drosophila imaginal wing disc. Genes Dev. 5, 996–1008.

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  2. Davis, C. A., Holmyard, D. P., Millen, K. J., and Joyner, A. L. (1991) Examining pattern formation in mouse, chicken and frog embryos with an En-specific antiserum. Development 111, 287–298.

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  3. Conlon, R. A. and Rossant, J. (1992) Exogenous retinoic acid rapidly induces anterior ectopic expression of murine Hox-2 genes in vivo. Development 116, 357–368.

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© 1997 Humana Press Inc., Totowa, NJs

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Conlon, R.A. (1997). Methods for Double Detection of Gene Expression. 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:271

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  • DOI: https://doi.org/10.1385/0-89603-481-X:271

  • 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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