Hostname: page-component-788cddb947-t9bwh Total loading time: 0 Render date: 2024-10-13T20:07:59.644Z Has data issue: false hasContentIssue false
  • English
  • Français

A Nonradiometric Detection System For the Rapid and Easy Detection of Oxytocin Oligonucleotide Probes in In Situ Hybridization

Published online by Cambridge University Press:  02 July 2020

M.C. Tyler  and
P.C. Mayer
M.C. Tyler
Affiliation:
NEN Life Science Products, 549 Albany Street, Boston, MA02118.
P.C. Mayer
Affiliation:
NEN Life Science Products, 549 Albany Street, Boston, MA02118.
Get access

Extract

We describe the easy and rapid detection of oxytocin oligonucleotide probes in in situ hybridization using a biotinyl tyramide amplification system (TSA).

There are several some instances where related proteins have sequence homology so great that only oligonucleotide probes will allow one to distinguish between their mRNAs. Vasopressin and oxytocin are such a pair. Traditional in situ hybridizationusing 35S labeled probes allows the specific detection of oxytocin mRNA.See FIG. 1.

Nonradiometnc detection of oligonucleotide probes can be very difficult since the amount of labeling possible is limited by the small size of the probe. Traditional nonradiometric detection of a 30-mer biotin-labeled oxytocin probe with streptavidin-HRP followed by DAB detection proved essentially negligible. See FIG. 2.

A novel tyramide amplification system (TSA) was used to increase the detection of the oxytocin probe signal. Most of the protocol was unchanged from the standard nonradioactive methodology. The biotin-labeled probe was hybridized overnight as usual. Stringency washes were also done as usual the next morning.

Type
Cytochemistry, Histochemistry, Immunocytochemistry, and In Situ Hybridization
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 145 - 146
Copyright
Copyright © Microscopy Society of America 1997

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1.Ivell, R., et al., Proc. Natl. Acad. Sci USA 81(1984)2006.10.1073/pnas.81.7.2006CrossRefGoogle Scholar
2.Young, W.S., et al., Mol. Brain Res. 1 (1986)231.10.1016/0169-328X(86)90029-XCrossRefGoogle Scholar
3.Burback, J.P.H., et al., Biochem. Biophys. Res. Comm. 145(1987)10.10.1016/0006-291X(87)91280-0CrossRefGoogle Scholar
4.Bobrow, M.N., et al., J. Immunol. Methods 125(1989)279.10.1016/0022-1759(89)90104-XCrossRefGoogle Scholar