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Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures
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Neurociência
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JoVE Journal Neurociência
Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures

Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures

DOI:
10.3791/50273-v

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13:50 min

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April 03, 2013

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, , ,
1Department of Cell Biology, Neurobiology and Anatomy,Medical College of Wisconsin , 2Department of Microbiology and Molecular Genetics,Medical College of Wisconsin

Capítulos

  • 00:05Título
  • 02:03Fixation
  • 05:31Immunohistochemistry
  • 11:30Results: Distribution of Endogenous Ng Molecules in Dendritic Spines
  • 13:03Conclusion

Summary

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The localization and distribution of proteins provide important information for understanding their cellular functions. The superior spatial resolution of electron microscopy (EM) can be used to determine the subcellular localization of a given antigen following immunohistochemistry. For tissues of the central nervous system (CNS), preserving structural integrity while maintaining antigenicity has been especially difficult in EM studies. Here, we adopt a procedure that has been used to preserve structures and antigens in the CNS to study and characterize synaptic proteins in rat hippocampal CA1 pyramidal neurons.

Tags

Immunogold LabelingPost-embeddingSynaptic ProteinsHippocampal Slice CulturesElectron MicroscopyUltrastructural LocalizationOsmium TetroxideUranyl AcetateTannic AcidAntigen DetectionStructural Preservation

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