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Curr Protoc Protein Sci. 2015 Feb 2;79:19.25.1-26. doi: 10.1002/0471140864.ps1925s79.

Measuring protein-protein and protein-nucleic Acid interactions by biolayer interferometry.

Author information

1
Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada.

Abstract

Biolayer interferometry (BLI) is a simple, optical dip-and-read system useful for measuring interactions between proteins, peptides, nucleic acids, small molecules, and/or lipids in real time. In BLI, a biomolecular bait is immobilized on a matrix at the tip of a fiber-optic sensor. The binding between the immobilized ligand and another molecule in an analyte solution produces a change in optical thickness at the tip and results in a wavelength shift proportional to binding. BLI provides direct binding affinities and rates of association and dissociation. This unit describes an efficient approach using streptavidin-based BLI to analyze DNA-protein and protein-protein interactions. A quantitative set of equilibrium binding affinities (K(d)) and rates of association and dissociation (k(a)/k(d)) can be measured in minutes using nanomole quantities of sample.

KEYWORDS:

binding affinities; biolayer interferometry; kinetic; protein-DNA interaction; protein-protein interaction

PMID:
25640894
DOI:
10.1002/0471140864.ps1925s79
[Indexed for MEDLINE]

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