Affinity Purification of FLAG-tagged hEAG1 Channel Protein from HEK-293 Cells
4:18
BCA Assay, Western Blotting, and Labeling the Purified Protein with Biotin for the BLI Assay
6:10
Bio-layer Interferometry Assay
8:06
Results: Interaction Kinetics of hEAG1 with PIP2 Using the Bio-layer Interferometry Assay
9:35
Conclusion
TRANSCRIÇÃO
The overall goal of this bio-layer interferometry or BLI assay, is to measure the potential interaction between purified hEAG1 ion channel protein and small molecule lipids. This method can help answer key questions in the ion channel pharmacology
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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.