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.
1. Biosensor hydration and assay set-up
2. Loading of ligand onto biosensor
3. Washing away additional ligand
4. Association of analyte to ligand
5. Dissociation of analyte from ligand
The authors have nothing to disclose.
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 |