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- W4379113362 abstract "Biolayer interferometry (BLI) is a widely utilized technique for determining the interaction dynamics between macromolecules. Most BLI instruments, such as the Octet RED96e used throughout this protocol, are completely automated and detect changes in the interference pattern of white light reflected off a biosensor tip. Biosensors are initially loaded with a stationary macromolecule, then introduced into a solution containing macromolecules of interest. Binding to the stationary molecules creates a change in optical wavelength that is recorded by the instrument in real-time. The majority of published BLI experiments assess protein-protein (such as antibody-substrate kinetics) or protein-small molecule (such as drug discovery) interactions. However, a less-appreciated assay for BLI analysis is DNA-protein interactions. In our laboratory, we have shown the practicality of using biotinylated-DNA probes to determine the binding kinetics of transcription factors to specific DNA sequences. The following protocol describes these steps, including the generation of biotinylated-DNA probes, the execution of the BLI experiment, and data analysis through GraphPad Prism. (The last step in this version contains a supplemental video with extra context and tips, as part of the protocols.io Spotlight series, featuring conversations with protocol authors.)" @default.
- W4379113362 created "2023-06-03" @default.
- W4379113362 creator A5000431436 @default.
- W4379113362 date "2023-06-02" @default.
- W4379113362 modified "2023-09-29" @default.
- W4379113362 title "Biolayer Interferometry for DNA-Protein Interactions v2" @default.
- W4379113362 doi "https://doi.org/10.17504/protocols.io.36wgq48movk5/v2" @default.
- W4379113362 hasPublicationYear "2023" @default.
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