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Title: Imaging beads-retained prey assay for rapid and quantitative protein-protein interaction
Authors: Zhou, Y.
Hong, Wanjin
Lu, Lei
Keywords: DRNTU::Science::Biological sciences
Issue Date: 2013
Source: Zhou, Y., Hong, W., & Lu, L. (2013). Imaging Beads-Retained Prey Assay for Rapid and Quantitative Protein-Protein Interaction. PLoS ONE, 8(3).
Series/Report no.: PLoS ONE
Abstract: Conventional Western blot based pull-down methods involve lengthy and laborious work and the results are generally not quantitative. Here, we report the imaging beads-retained prey (IBRP) assay that is rapid and quantitative in studying protein-protein interactions. In this assay, the bait is immobilized onto beads and the prey is fused with a fluorescence protein. The assay takes advantage of the fluorescence of prey and directly quantifies the amount of prey binding to the immobilized bait under a microscope. We validated the assay using previously well studied interactions and found that the amount of prey retained on beads could have a relative linear relationship to both the inputs of bait and prey. IBRP assay provides a universal, fast, quantitative and economical method to study protein interactions and it could be developed to a medium- or high-throughput compatible method. With the availability of fluorescence tagged whole genome ORFs in several organisms, we predict IBRP assay should have wide applications.
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0059727
Schools: School of Biological Sciences 
Rights: © 2013 Zhou et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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