SUBCELLULAR-LOCALIZATION OF ALPHA-TUBULIN AND OPSIN MESSENGER-RNA IN THE GOLDFISH RETINA USING DIGOXIGENIN-LABELED CRNA PROBES DETECTED BY ALKALINE-PHOSPHATASE AND HRP HISTOCHEMISTRY

Citation
Lk. Barthel et Pa. Raymond, SUBCELLULAR-LOCALIZATION OF ALPHA-TUBULIN AND OPSIN MESSENGER-RNA IN THE GOLDFISH RETINA USING DIGOXIGENIN-LABELED CRNA PROBES DETECTED BY ALKALINE-PHOSPHATASE AND HRP HISTOCHEMISTRY, Journal of neuroscience methods, 50(2), 1993, pp. 145-152
Citations number
26
Categorie Soggetti
Neurosciences
ISSN journal
01650270
Volume
50
Issue
2
Year of publication
1993
Pages
145 - 152
Database
ISI
SICI code
0165-0270(1993)50:2<145:SOAAOM>2.0.ZU;2-3
Abstract
This paper describes a method for non-radioactive in situ hybridizatio n providing subcellular localization of mRNA in 3 mu m cryosections. W e used two alternative colorimetric reactions to detect digoxigenin-la beled cRNA probes: alkaline phosphatase and HRP (horseradish peroxidas e). With some probes the signal with the alkaline phosphatase reaction was intense, and diffusion of the reaction product was noticeable. Us ing HRP-conjugated antibodies improved the resolution but decreased th e sensitivity of the signal. Photoamplification of the HRP reaction pr oduct increased the contrast and improved the sensitivity of the techn ique.