title: A Monoclonal Antibody That Discriminates Between SNAP-Tagged and CLIP-Tagged Proteins.
abstract: SNAP-tag technology allows recombinant proteins to be covalently labeled to O(6)-benzylguanine (BG)-modified substrates with 1:1 stoichiometry. By attaching according fluorophores, this method is ideally suited for in vitro and in vivo imaging, as well as protein interaction analyses. Fluorophores modified with BG react with the SNAP-tag, whereas those modified with O(2)-benzylcytosine (BC) conjugate to a more recent derivative known as the CLIP-tag. The orthogonal substrate specificity of the SNAP- and CLIP-tags extends the range of applications by allowing double labeling. We previously developed a monoclonal antibody (mAb) that recognizes both tags. In this study, we describe a new mAb, which is specific for the SNAP-tag alone. Therefore, this mAb allows discrimination between SNAP- and CLIP-tags within a broad range of immunological methods, including enzyme-linked immunosorbent assays, western blotting, flow cytometry, and immunohistochemistry.
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .
authors.forename: Magdalena
authors.initials: M
authors.lastname: Bialon
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .|2 Department of Molecular Biotechnology, RWTH Aachen University , Aachen, Germany .
authors.forename: Clara
authors.initials: C
authors.lastname: Grezella
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .
authors.forename: Ludmila
authors.initials: L
authors.lastname: Friesen
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .
authors.forename: Thorsten
authors.initials: T
authors.lastname: Sieben
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .
authors.forename: Anh-Tuan
authors.initials: AT
authors.lastname: Pham
authors.affiliation: 2 Department of Molecular Biotechnology, RWTH Aachen University , Aachen, Germany .|3 Fraunhofer Institute for Molecular Biology and Applied Ecology , Aachen, Germany .
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .|3 Fraunhofer Institute for Molecular Biology and Applied Ecology , Aachen, Germany .
authors.forename: Stefan
authors.initials: S
authors.lastname: Barth
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .
authors.forename: Christiane
authors.initials: C
authors.lastname: Püttmann
authors.affiliation: 1 Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, Institute of Applied Medical Engineering, University Hospital RWTH Aachen , Aachen, Germany .|3 Fraunhofer Institute for Molecular Biology and Applied Ecology , Aachen, Germany .
authors.forename: Christoph
authors.initials: C
authors.lastname: Stein
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D006147:Guanine / Q000031:analogs & derivatives* / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; C064976:O(6)-benzylguanine; D006147:Guanine