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
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.