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Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay
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Biochemistry
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JoVE Journal Biochemistry
Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay

Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay

DOI:
10.3791/56846-v

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10:41 min

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March 07, 2018

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, , ,
1Department of General Surgery, Tongren Hospital,Shanghai Jiao Tong University School of Medicine, 2Key Laboratory of Systems Biomedicine (Ministry of Education), Institute of Systems Biomedicine,Shanghai Jiao Tong University, 3Hongqiao International Institute of Medicine, Tongren Hospital,Shanghai Jiao Tong University School of Medicine

Chapters

  • 00:04Title
  • 01:01Affinity Purification of FLAG-tagged hEAG1 Channel Protein from HEK-293 Cells
  • 04:18BCA Assay, Western Blotting, and Labeling the Purified Protein with Biotin for the BLI Assay
  • 06:10Bio-layer Interferometry Assay
  • 08:06Results: Interaction Kinetics of hEAG1 with PIP2 Using the Bio-layer Interferometry Assay
  • 09:35Conclusion

Summary

Automatic Translation

Automatic Translation

The protocol here describes the interactions of purified hEAG1 ion channel protein with the small molecule lipid ligand phosphatidylinositol 4, 5-bisphosphate (PIP2). The measurement demonstrates that BLI could be a potential method for novel small-molecule ion channel ligand screening.

Tags

Keywords: Bio-layer InterferometryBLI AssayIon ChannelProtein-small Molecule InteractionKineticsHEAG1Cell CultureProtein ExpressionPurificationAffinity Chromatography

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