Detection of the human GPR50 orphan seven transmembrane protein by polyclonal antibodies mapping different epitopes
Autor: | Hassina Ould Hamouda, Pascal Maurice, Ralf Jockers, Nejla Sözer-Topçular, Jean-Luc Guillaume, Avais M. Daulat, Gilles Ferry, Jean A. Boutin, Angélique Levoye, Rivka Ravid, Patty Chen, Egemen Savaskan, Philippe Delagrange |
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Přispěvatelé: | Institut Cochin (UMR_S567 / UMR 8104), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), University of Basel (Unibas), Netherlands Brain Bank [Amsterdam, The Netherlands] (N2B), Institut de Recherches Internationales Servier [Suresnes] (IRIS), Centre de Recherche en Cancérologie de Marseille (CRCM), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherches Servier, Centre de Recherches de Croissy, Département des Sciences Expérimentales, Institut de Recherches Internationales Servier, University of Zurich, Jockers, R, MAURICE, Pascal |
Jazyk: | angličtina |
Rok vydání: | 2007 |
Předmět: |
[SDV]Life Sciences [q-bio]
Molecular Sequence Data Nerve Tissue Proteins 610 Medicine & health CHO Cells Immunofluorescence Antibodies Epitope Cell Line Receptors G-Protein-Coupled Epitopes 03 medical and health sciences Cricetulus 0302 clinical medicine Endocrinology Western blot Sequence Analysis Protein Cricetinae medicine Animals Humans Amino Acid Sequence ComputingMilieux_MISCELLANEOUS 030304 developmental biology Orphan receptor 0303 health sciences biology medicine.diagnostic_test 11359 Institute for Regenerative Medicine (IREM) Fusion protein Molecular biology Transmembrane protein Protein Structure Tertiary 1310 Endocrinology [SDV] Life Sciences [q-bio] Epitope mapping Polyclonal antibodies biology.protein Binding Sites Antibody Rabbits Epitope Mapping 030217 neurology & neurosurgery |
Zdroj: | Journal of Pineal Research Journal of Pineal Research, Wiley, 2007, 43 (1), pp.10-15. ⟨10.1111/j.1600-079X.2007.00437.x⟩ Journal of Pineal Research, 2007, 43 (1), pp.10-15. ⟨10.1111/j.1600-079X.2007.00437.x⟩ |
ISSN: | 0742-3098 1600-079X |
DOI: | 10.1111/j.1600-079X.2007.00437.x⟩ |
Popis: | International audience; GPR50 is an orphan seven transmembrane protein related to the melatonin receptor subfamily comprising MT(1) and MT(2) receptors. In the absence of any known ligand for GPR50, other tools are critical for the characterization of this protein. Here, we describe the generation, purification and characterization of the first rabbit polyclonal antibodies generated against peptides corresponding to the N-terminus, C-terminus and two additional regions within the intracellular tail of GPR50. Immune sera were purified on peptide-antigen affinity columns. Antibodies specifically recognized a GPR50-YFP fusion protein on the plasma membrane of HEK 293 cells in immunofluorescence experiments. In Western blot experiments, the monomeric and dimeric forms of GPR50 were detected as proteins of 66 and 130 kDa, respectively. In addition, these new antibodies were sufficiently sensitive to detect GPR50 in brain slices of the rat pituitary and human hippocampus. In conclusion, we successfully produced antibodies against the orphan GPR50 protein that will become valuable tools for functional studies of this protein. |
Databáze: | OpenAIRE |
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