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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.
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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

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References

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