Journal
/
/
Påvisning af protein palmitoylering i dyrkede hippocampale neuroner ved immunopræcipitation og Acyl-Biotin Exchange (ABE)
Journal JoVE
Neurosciences
This content is Free Access.
Journal JoVE Neurosciences
Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange (ABE)
DOI:

16:33 min

February 18, 2013

,

Chapitres

  • 00:05Titre
  • 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

Traduction automatique

Den reversible tilsætning af palmitat til proteiner er en vigtig regulator af intracellulært protein trafficking. Dette er af særlig interesse i neuroner, hvor mange synaptiske proteiner palmitoylated. Vi anvender en enkel biokemisk metode til at påvise palmitoylated proteiner i dyrkede neuroner, som kan tilpasses til flere celletyper og væv.

Vidéos Connexes

Read Article