title: Monoclonal Antibodies 13A4 and AC133 Do Not Recognize the Canine Ortholog of Mouse and Human Stem Cell Antigen Prominin-1 (CD133).
abstract: The pentaspan membrane glycoprotein prominin-1 (CD133) is widely used in medicine as a cell surface marker of stem and cancer stem cells. It has opened new avenues in stem cell-based regenerative therapy and oncology. This molecule is largely used with human samples or the mouse model, and consequently most biological tools including antibodies are directed against human and murine prominin-1. Although the general structure of prominin-1 including its membrane topology is conserved throughout the animal kingdom, its primary sequence is poorly conserved. Thus, it is unclear if anti-human and -mouse prominin-1 antibodies cross-react with their orthologs in other species, especially dog. Answering this issue is imperative in light of the growing number of studies using canine prominin-1 as an antigenic marker. Here, we address this issue by cloning the canine prominin-1 and use its overexpression as a green fluorescent protein fusion protein in Madin-Darby canine kidney cells to determine its immunoreactivity with antibodies against human or mouse prominin-1. We used immunocytochemistry, flow cytometry and immunoblotting techniques and surprisingly found no cross-species immunoreactivity. These results raise some caution in data interpretation when anti-prominin-1 antibodies are used in interspecies studies.
authors.affiliation: Tissue Engineering Laboratories, Biotechnology Center (BIOTEC), Technische Universität Dresden, Dresden, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Thamm
authors.affiliation: Tissue Engineering Laboratories, Biotechnology Center (BIOTEC), Technische Universität Dresden, Dresden, Germany.
authors.forename: Sylvi
authors.initials: S
authors.lastname: Graupner
authors.affiliation: DFG-Research Center and Cluster of Excellence for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Dresden, Germany.|Institute for Biofunctional Polymer Materials, Leibniz Institute of Polymer Research Dresden, Dresden, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Werner
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
authors.forename: Wieland B
authors.initials: WB
authors.lastname: Huttner
authors.affiliation: Tissue Engineering Laboratories, Biotechnology Center (BIOTEC), Technische Universität Dresden, Dresden, Germany.|DFG-Research Center and Cluster of Excellence for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Dresden, Germany.
authors.forename: Denis
authors.initials: D
authors.lastname: Corbeil
mesh_terms: D000071916:AC133 Antigen / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D018938:Caco-2 Cells; D003001:Cloning, Molecular / Q000379:methods*; D004285:Dogs; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D013045:Species Specificity
chemical_list: D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal