Two-color detection of mRNA transcript localizations in fish and fly embryos using alkaline phosphatase and beta-galactosidase conjugated antibodies

Authors
Citation
G. Hauptmann, Two-color detection of mRNA transcript localizations in fish and fly embryos using alkaline phosphatase and beta-galactosidase conjugated antibodies, DEV GENES E, 209(5), 1999, pp. 317-321
Citations number
21
Categorie Soggetti
Cell & Developmental Biology
Journal title
DEVELOPMENT GENES AND EVOLUTION
ISSN journal
0949944X → ACNP
Volume
209
Issue
5
Year of publication
1999
Pages
317 - 321
Database
ISI
SICI code
0949-944X(199905)209:5<317:TDOMTL>2.0.ZU;2-P
Abstract
A fast and simplified two-color in situ hybridization procedure for the sim ultaneous detection of two different mRNAs in whole-mounted zebrafish and D rosophila embryos is presented. Transcript distributions are detected in a single incubation step using a mixture of alkaline phosphatase and beta-gal actosidase coupled antibodies. The different transcripts are visualized in contrasting colors by the use of beta-galactosidase substrates that develop color precipitates (magenta, blue) easily distinguishable from those of th e standard alkaline phosphatase substrates. This protocol can be followed b y standard immunohistochemistry to detect the expression of a third gene (a t the protein level) in a third color.