title: In Situ Protein Binding Assay Using Fc-Fusion Proteins.
abstract: This protocol describes an in situ protein-protein interaction assay between tagged recombinant proteins and cell-surface expressed synaptic proteins. The assay is arguably more sensitive than other traditional protein binding assays such as co-immunoprecipitation and pull-downs and provides a visual readout for binding. This assay has been widely used to determine the dissociation constant of binding of trans-synaptic adhesion proteins. The step-wise description in the protocol should facilitate the adoption of this method in other laboratories.
authors.affiliation: Department of Physiology and Pathophysiology, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada.|Neuroscience Research Program, Kleysen Institute for Advanced Medicine, Health Sciences Centre, University of Manitoba, 710 William Avenue, Winnipeg, MB, Canada, R3E 0Z3.
authors.forename: Nirmala
authors.initials: N
authors.lastname: Padmanabhan
authors.affiliation: Department of Physiology and Pathophysiology, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada. tabrez.siddiqui@umanitoba.ca.|Neuroscience Research Program, Kleysen Institute for Advanced Medicine, Health Sciences Centre, University of Manitoba, 710 William Avenue, Winnipeg, MB, Canada, R3E 0Z3. tabrez.siddiqui@umanitoba.ca.
authors.forename: Tabrez J
authors.initials: TJ
authors.lastname: Siddiqui
mesh_terms: D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000378:metabolism*; D018929:Cell Culture Techniques; D002522:Chlorocebus aethiops; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins
keywords: Fc-fusion protein; In situ protein binding assay; Protein-protein interaction