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Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions
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Biochemistry
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JoVE Journal Biochemistry
Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions

Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions

DOI:
10.3791/56457-v

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10:26 min

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December 20, 2017

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, , , ,
1Instituto de Neurociencias de Castilla y León (INCYL), Departamento de Bioquímica y Biología Molecular,Universidad de Salamanca, 2Centre for Cancer Research & Cell Biology (CCRCB), School of Medicine, Dentistry and Biomedical Sciences,Queen’s University Belfast

Chapters

  • 00:05Title
  • 00:45Cells and Biotinylated CPPs
  • 02:01Cellular Treatment with the BCPPs
  • 03:49Protein Extraction and Pull-down
  • 07:19Results: Study of the Intracellular TAT-Cx43266-283-B Interactions in G166 GSCs
  • 09:00Conclusion

Summary

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This is a protocol to study intracellular protein-protein interactions based on the biotin-avidin pull-down system with the novelty of combining cell-penetrating sequences. The main advantage is that the target sequence is incubated with living cells instead of cell lysates and therefore the interactions will occur within the cellular context.

Tags

Keywords: Biotinylated Cell-penetrating PeptidesProtein-protein InteractionsIntracellular ContextBiotin-avidin Pull-down SystemSelf-penetrating SequencesCell SignalingG166 Glioma Stem CellsGSCsBCPPNeutrAvidinWestern Blot

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