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Påvisning av Protein palmitoylation i Cultured hippocampus neurons ved Immunoutfellingsanalyse og Acyl-Biotin Exchange (ABE)
JoVE Journal
Neuroscience
This content is Free Access.
JoVE Journal Neuroscience
Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange (ABE)
DOI:

16:33 min

February 18, 2013

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Chapters

  • 00:05Title
  • 02:08Extraction of Target Protein from Cultured Primary Hippocampal Neurons
  • 05:54Acyl-Biotin Exchange: Hydroxylamine (HAM) Cleavage
  • 09:17Elution of Biotin-conjugated Target Protein and SDS-PAGE
  • 10:46Western Blotting and Detection of Palmitoylation of Target Protein
  • 12:16Results: Detecting Palmitoylation of a Neuronal Protein
  • 16:07Conclusion

Summary

Automatic Translation

Den reversible tilsetning av palmitate til proteiner er en viktig regulator av intracellulær protein menneskehandel. Dette er av spesiell interesse i nevroner hvor mange synaptiske proteiner palmitoylated. Vi benytter en enkel biokjemisk metode for å detektere palmitoylated proteiner i dyrkede nevroner, som kan tilpasses for flere celletyper og vev.

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