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Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections
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Neuroscience
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JoVE Journal Neuroscience
Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections

Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections

DOI:
10.3791/53976-v

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09:23 min

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March 26, 2016

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, , , ,
1Wolfson Institute for Biomedical Research,University College London

Chapters

  • 00:05Title
  • 00:42Overview of Double Fluoresence In Situ Hybridization Followed by Immunostaining
  • 01:44RNA Probe Synthesis
  • 04:12Cryosectioning and Hybridization
  • 05:33Visualization of the FITC Probe and the DIG Probe
  • 07:23Immunohistochemistry and Mounting
  • 08:16Results: Expression of Aspa in Oligodendrocytes in the Cortex and the Corpus Callosum
  • 08:45Conclusion

Summary

Automatic Translation

Automatic Translation

Localizing gene expression to specific cell types can be challenging due to the lack of specific antibodies. Here we describe a protocol for simultaneous triple detection of gene expression by combining double fluorescence RNA in situ hybridization with immunostaining.

Tags

Keywords: Double Fluorescence In Situ HybridizationImmunolabellingGene ExpressionMouse Brain SectionsCell Type LocalizationRNA ProbesFITCDIGTyramideAlkaline PhosphataseFast RedImmunostainingPlasmid DNARNA Polymerase PromotersCDNARestriction EnzymeLinearizationPurificationPhenol-chloroform ExtractionEthanol Precipitation

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