title: Measuring protein-protein and protein-nucleic Acid interactions by biolayer interferometry.
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.
authors: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada.
authors: Azmiri
authors: A
authors: Sultana
authors: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada.
authors: Jeffrey E
authors: JE
authors: Lee
mesh_terms: D001681:Biological Assay / Q000379:methods*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D007368:Interferometry / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D011506:Proteins; D004247:DNA
keywords: binding affinities; biolayer interferometry; kinetic; protein-DNA interaction; protein-protein interaction