Biolayer Interferometry Technology to Detect Interactions between Ligand and Analyte

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

1. Turn the BLItz machine on.
2. Ensure that the machine is connected to the computer through a USB data output port at the back of the machine.
3. On the computer, open the associated software (e.g., BLItz Pro), and click on Advanced Kinetics on the left-hand side of the screen.
4. On the software, type out all appropriate information about the experiment (including the Experiment Name, Description, Sample ID, and Protein Concentration) under each respective heading.
5. Click on Biosensor Type and choose Ni-NTA from the drop-down menu.
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Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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