By: Paul A Mould1 

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Solid-phase assays provide a simple, rapid and robust method for the analysis of protein–protein interactions; i.e., does
protein A interacts with protein B? In this assay, protein A (here termed as ‘receptor’) is adsorbed to the wells of an enzyme-linked
immunosorbent assay (ELISA) plate (solid phase). The plate is then blocked using bovine serum albumin (BSA), and biotin-labelled
protein B (here termed as ‘ligand’) is added. After washing the wells to remove unbound ligand, bound ligand is detected by
addition of an avidin–peroxidase conjugate followed by a colorimetric detection step. This type of assay is particularly well
suited for studying the interaction of ECM proteins with integrins. The screening of antagonists of integrin–ligand interactions
in the pharmaceutical industry is an important area in which this assay is finding use.
Affiliation(s): (1) B3070 Michael Smith Building, Wellcome Trust Centre for Cell-Matrix research, Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester, M13 9PT, UK
Book Title: Extracellular Matrix Protocols: Second Edition
Series: Methods in Molecular Biology | Volume: 522 | Pub. Date: Jun-01-2008 | Page Range: 1-6 | DOI: 10.1007/978-1-59745-413-1_13
Subject: Cell Biology
Key Words: Protein–protein interaction - Solid phase assay - Extracellular matrix - Integrin - Ligand binding - Divalent cations - Antagonists - Pharmacology
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