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Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
JoVE Journal
Biology
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JoVE Journal Biology
Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
DOI:

11:16 min

July 12, 2014

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Chapters

  • 00:05Title
  • 01:23Protein Pulldowns: Preparation of Transiently Transfected Cells
  • 02:50Equilibration of the HaloLink Resin
  • 04:01Binding and Washing of Fusion Complexes
  • 06:21SDS Elution for Denaturing Gels, Western Blots, or Mass Spectrometry
  • 07:07Cellular Imaging of Fluorescently Labeled Fusion Proteins Using a Cytoplasmic and Nuclear Permeable Ligand
  • 08:48Results: Using HaloTag Technology to Determine Protein Interactions and Localization
  • 10:47Conclusion

Summary

Automatic Translation

HaloTag technology is a multifunctional technology which has shown significant success in isolation of both small and large protein complexes from mammalian cells.  Here we highlight the advantages of this technology compared to existing alternatives and demonstrate its utility to study numerous aspects of protein function inside eukaryotic cells.

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