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Start Over Author "Bernard Liu" Journal methods in molecular biology (clifton, n.j.)
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1. Binding Assays Using Recombinant SH2 Domains: Far-Western, Pull-Down, and Fluorescence Polarization

Author

Kazuya, Machida and Bernard, Liu

Subjects
src Homology Domains, Recombinant Fusion Proteins, Blotting, Western, Luminescent Measurements, Fluorescent Antibody Technique, Fluorescence Polarization, Phosphorylation, Phosphotyrosine, Protein Binding
Abstract

Recognition of phosphotyrosine-containing sequences by SH2 domains confers specificity in tyrosine kinase pathways. By assessing interactions between isolated SH2 domains and their binding proteins, it is possible to gain insight into otherwise inaccessible complex cellular systems. Far-Western, pull-down, and fluorescence polarization (FP) have been frequently used for characterization of phosphotyrosine signaling. Here, we outline standard protocols for these established assays using recombinant SH2 domain, emphasizing the importance of appropriate sample preparation and assay controls.

Published
2017

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