Efficient Target Retrieval for the RNAscope Assay

July 30, 2026

BlueskyReddit
State University of New York at Buffalo
Physiology and Biophysics
Postdoctoral Researcher

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Company:

Advanced Cell Diagnostics / Bio-Techne

Product Name:

RNAscope™ Target Retrieval Reagents

Catalog Number:

322000

We utilize RNAscope combined with immunostaining to assess Cnr1 mRNA (the Cnr1 gene encodes cannabinoid receptor 1) in astrocytes of mouse brain tissue. RNAscope Target Retrieval Reagent increases the accessibility of RNA molecules and unmasks antigenic epitopes that become hidden during fixation, allowing our RNAscope assay to work consistently well.

Experimental Design and Results Summary

Application

RNAscope combined with immunostaining

Starting Material

15 μm fixed-frozen mouse brain sections

Tips

We follow the manufacturer’s instructions (User Manual UM 323100/Rev C, ACD) for the use of target retrieval during the RNAscope assay. We use the “hot plate method” and place the samples (slides) into a beaker containing Target Retrieval Reagent heated to 98–102°C. Before placing the samples into the Target Retrieval Reagent, we briefly submerge the slides in mildly boiling distilled H₂O for 10 seconds to acclimate them. A 5-minute boiling step was optimal for our samples (15 μm fixed-frozen brain tissue sections).

Results Summary

They were able to assess Cnr1 mRNA expression (red dots) in astrocytes (green, immunostained for S100β) in mouse dorsal striatum and cortex tissue. We compared astrocytic expression of cannabinoid receptors among mice exposed to adverse environmental factors and control mice.

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Summary

The Good

The reagent comes in four bottles, which are sufficient for four experiments. It also has a good shelf life of two years.

The Bad

We did not observe any adverse effects with this reagent. It worked as expected.

The Bottom Line

The reagent is convenient to use (it is supplied as a concentrate that must be diluted in water according to the manufacturer’s instructions) and provides reliable results. It improves signal intensity while maintaining low background. Highly recommended.

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