ABSTRACT
Biotin-UTP or digoxigenin-UTP are now commonly used in the synthesis of nonradiolabeled nucleic acid probes.
The advantages for employing nonradiolabeled probes for
in situ
hybridization studies are the improved stability of the nonradiolabeled probe, greater cellular resolution for the hybridization signal, an improved signal-to-noise ratio, and the decreased time to observe the hybridization signal. With these advantages, nonradiolabeled probes have been more widely employed in recent years. For example, digoxigenin (DIG)-labeled probes have been used to localize expression of keratin 17,
type I hair keratin genes,
involucrin,
fibroblast growth factor receptor and ligand genes,
and mouse Notch
to skin and hair follicles. Thus, it also seems appropriate to review methods for ISH using a nonradiola-
beled probe; in particular, the use of DIG-labeled riboprobes for
in situ
hybridization will be covered. A standard detection kit is readily available from BoehringerMannheim (the Genius
Nucleic Acid Labeling and Detection Kit, Indianapolis, IN). Boehringer-Mannheim also has developed a
Non-radioactive In Situ Hybridization Application Manual
(Second Edition) to review applications for DIG-labeled DNA-or RNA-labeled probes.
