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用生物层干涉法捕获小分子离子通道蛋白的相互作用动力学
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Bioquímica
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JoVE Journal Bioquímica
Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay
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用生物层干涉法捕获小分子离子通道蛋白的相互作用动力学

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

Capítulos

  • 00:04Título
  • 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

Tadução automática

Tadução automática

该协议描述了纯化 hEAG1 离子通道蛋白与小分子脂配体磷酸肌醇4、5-bisphosphate (PIP2) 的相互作用。实验表明, BLI 可能是一种新的小分子离子通道配体筛选的潜在方法。

Tags

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

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