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Kombinere Double Fluorescence<em> In Situ</em> Hybridisering med immunomerking for Påvisning av Expression of Three Gener i musehjerne §§
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JoVE 杂志 神经科学
Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections

Kombinere Double Fluorescence<em> In Situ</em> Hybridisering med immunomerking for Påvisning av Expression of Three Gener i musehjerne §§

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

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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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