Tom I, et al. (2013) Protein microarrays for identification of novel extracellular protein-protein interactions. Curr Protoc Protein Sci Chapter 27():Unit 27.3
Abstract: Functional protein microarrays offer the capability for high-throughput protein interaction analysis and have long promised to be a powerful tool for understanding protein interactions at the proteome scale. Although popular techniques for protein-protein interaction mapping like yeast-two-hybrid and affinity-purification mass spectrometry have performed well for identifying intracellular protein-protein interactions, the study of interactions between extracellular proteins has remained challenging for these methods. Instead, the use of protein microarrays appears to be a robust and efficient method for the identification of interactions among the members of this class of protein. This unit describes methods for extracellular protein microarray production, screening, and analysis. A protocol is described for enhanced detection of low-affinity interactions by generating multivalent complexes using Fc-fusion bait proteins and protein A microbeads, along with a statistical method for hit scoring and identification of nonspecific interactions. Curr. Protoc. Protein Sci. 72:27.3.1-27.3.24. (c) 2013 by John Wiley & Sons, Inc.
|Status: Published||Type: Journal Article||PubMed ID: 23546621|