Biolayer Interferometry Technology to Detect Interactions between Ligand and Analyte
Published: May 31, 2023
Abstract
Source: Desai, M., et al. Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription. J. Vis. Exp. (2019)
In this video, we demonstrate biolayer interferometry (BLI) technology to detect the intermolecular interactions between ligands and target analytes in real-time. BLI measures changes in the interference pattern of white light reflected off a layer of immobilized ligands on a biosensor surface as they interact with analytes in solution.
Protocol
1. Biosensor hydration and assay set-up Turn the BLItz machine on. Ensure that the machine is connected to the computer through a USB data output port at the back of the machine. On the computer, open the associated software (e.g., BLItz Pro), and click on Advanced Kinetics on the left-hand side of the screen. On the software, type out all appropriate information about the experiment (including the Experiment Name, Description, Sample ID, and Protein Concentratio…
Offenlegungen
The authors have nothing to disclose.
Materials
| BLItz machine | ForteBio | 45-5000 | Device |
| Dip and Read Ni-NTA biosensor tray | ForteBio | 18-5101 | Ready-to-use Ni-NTA biosensors for poly-His-tagged Proteins |
| Drop holder | ForteBio | 45-5004 | Device |
| PCR tubes (0.2 mL) | Thomas Scientific | CLS6571 | Plastic ware |
| Microcentrifuge tubes (black) | Thermo Fisher Scientific | 03-391-166 | Plastic ware |
| Kimwipes | Thermo Fisher Scientific | 06-666A | Plastic ware |
| DTT | Thermo Fisher Scientific | R0861 | Chemical |
| EDTA | MilliporeSigma | E6758 | Chemical |
| MgCl2 | MilliporeSigma | M8266 | Chemical |
| NaCl | MilliporeSigma | S9888 | Chemical |
| Tris-HCl | GoldBio | T095100 | Buffer |
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Diesen Artikel zitieren
Biolayer Interferometry Technology to Detect Interactions between Ligand and Analyte. J. Vis. Exp. (Pending Publication), e21398, doi: (2023).