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Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks
JoVE Journal
Biology
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JoVE Journal Biology
Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

DOI:
10.3791/54870-v

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11:04 min

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December 24, 2016

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, , , ,
1Department of Chemistry and Biochemistry,University of California, Los Angeles, 2Molecular Biology Institute,University of California, Los Angeles, 3Jonsson Comprehensive Cancer Center,University of California, Los Angeles

Chapters

  • 00:05Title
  • 00:44Generation of an Inducible Stable Cell Line that Expresses the LAP-tagged Gene of Interest
  • 02:06Purification of LAP-tagged Protein Complexes
  • 08:10Identify Interacting Proteins by Mass Spectrometry Analysis
  • 09:12Results: Evaluation of a LAP-Tau HEK293 Inducible Cell Line
  • 10:32Conclusion

Summary

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Automatic Translation

We describe a method for generating localization and affinity purification (LAP)-tagged inducible stable cell lines for investigating protein function, spatiotemporal subcellular localization and protein-protein interaction networks.

Tags

Keywords: Inducible LAP-taggedStable Cell LinesProtein FunctionSpatiotemporal LocalizationProtein Interaction NetworksHEK293 CellsHygromycin SelectionTandem Affinity Purification (TAP)Tet/Dox InductionAnti-GFP AntibodyProtein A Beads

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