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Solid Phase Assays for Studying ECM Protein–Protein Interactions

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Extracellular Matrix Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 522))

Summary

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.

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References

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Correspondence to Paul A Mould .

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Mould, P. (2009). Solid Phase Assays for Studying ECM Protein–Protein Interactions. In: Even-Ram, S., Artym, V. (eds) Extracellular Matrix Protocols. Methods in Molecular Biology, vol 522. Humana Press. https://doi.org/10.1007/978-1-59745-413-1_13

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  • DOI: https://doi.org/10.1007/978-1-59745-413-1_13

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-984-0

  • Online ISBN: 978-1-59745-413-1

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