Abstract:
BACKGROUND:Analysis of protein-protein interactions (PPIs) is a valuable approach for the characterization of huge networks of protein complexes or proteins of unknown function. Co-immunoprecipitation (coIP) using affinity resins coupled to protein A/G is the most widely used method for PPI detection. However, this traditional large scale resin-based coIP is too laborious and time consuming. To overcome this problem, we developed a miniaturized sandwich immunoassay platform (MSIP) by combining antibody array technology and coIP methods. RESULTS:Based on anti-FLAG antibody spotted aldehyde slides, MSIP enables simple, rapid and large scale detection of PPIs by fluorescent labeling anti-myc antibody. By analyzing well-known interacting and non-interacting protein pairs, MSIP was demonstrated to be highly accurate and reproducible. Compared to traditional resin-based coIP, MSIP results in higher sensitivity and enhanced throughput, with the additional benefit of digital read-outs. In addition, MSIP was shown to be a highly useful validation platform to confirm PPI candidates that have been identified from yeast two hybrid systems. CONCLUSIONS:In conclusion, MSIP is proved to be a simple, cost-saving and highly efficient technique for the comprehensive study of PPIs.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Liu Q,Chen Q,Wang J,Zhang Y,Zhou Y,Lin C,He W,He F,Xu Ddoi
10.1186/1472-6750-10-78subject
Has Abstractpub_date
2010-10-28 00:00:00pages
78issn
1472-6750pii
1472-6750-10-78journal_volume
10pub_type
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