title: Far Western blotting as a rapid and efficient method for detecting interactions between DNA replication and DNA repair proteins.
abstract: Protein-protein interactions are required for the proper function of many biological pathways. Numerous biochemical and protein blotting methods are available for probing direct and indirect interactions between two protein-binding partners. Here, we describe the methodology of far Western blotting, or immunodot blotting, as a technique for probing direct interactions between two proteins. We describe the utility of this approach as a rapid, qualitative screen for identifying novel protein-binding partners. We also describe the importance of this technique for measuring differences in interaction between wild-type and mutant forms of a known binding partner. Far Western blotting is a rapid and highly reproducible experimental approach for identifying and understanding the interaction between protein-binding partners leading to new discoveries in the function and regulation of biological pathways.
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Brian W
authors.initials: BW
authors.lastname: Walsh
authors.forename: Justin S
authors.initials: JS
authors.lastname: Lenhart
authors.forename: Jeremy W
authors.initials: JW
authors.lastname: Schroeder
authors.forename: Lyle A
authors.initials: LA
authors.lastname: Simmons
mesh_terms: D026481:Blotting, Far-Western / Q000379:methods; D004260:DNA Repair; D004261:DNA Replication; D008967:Molecular Biology / Q000379:methods; D051719:MutS DNA Mismatch-Binding Protein / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D013997:Time Factors
chemical_list: D051719:MutS DNA Mismatch-Binding Protein