title: Subunit-specific polyclonal antibody targeting human ρ1 GABA(C) receptor.
abstract: The GABA(C) receptor, a postsynaptic membrane receptor expressed prominently in the retina, is a ligand-gated ion channel that consists of a combination of ρ subunits. We report characterization of a novel guinea pig polyclonal antibody, termed GABA(C) Ab N-14, directed against a 14-mer peptide (N-14) of the extracellular domain of the human ρ1 subunit. The antibody exhibits high sensitivity for N-14 by ELISA. In Western blots, GABA(C) Ab N-14 shows reactivity with the ρ1 subunit of preparations obtained from ρ1 GABA(C)-expressing neuroblastoma cells, Xenopus oocytes, and mammalian retina and brain. Flow cytometry reveals a rightward shift in mean fluorescence intensity of GABA(C)-expressing neuroblastoma cells probed with GABA(C) Ab N-14. Immunostaining of neuroblastoma cells and oocytes with GABA(C) Ab N-14 yields fluorescence only with GABA(C)-expressing cells. Antibody binding has no effect on GABA-elicited membrane current responses. Immunostaining of human retinal sections shows preferential staining within the inner plexiform layer. GABA(C) Ab N-14 appears well suited for investigative studies of GABA(C) ρ1 subunit in retina and other neural tissues.
authors.affiliation: Lions of Illinois Eye Research Institute, Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, 1855 W. Taylor St., Chicago, IL 60612, USA.
authors.forename: Hélène A
authors.initials: HA
authors.lastname: Gussin
authors.forename: Fadi T
authors.initials: FT
authors.lastname: Khasawneh
authors.forename: An
authors.initials: A
authors.lastname: Xie
authors.forename: Feng
authors.initials: F
authors.lastname: Feng
authors.forename: Adnan
authors.initials: A
authors.lastname: Memic
authors.forename: Haohua
authors.initials: H
authors.lastname: Qian
authors.forename: Guy C
authors.initials: GC
authors.lastname: Le Breton
authors.forename: David R
authors.initials: DR
authors.lastname: Pepperberg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000097:blood*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D001921:Brain / Q000276:immunology; D002846:Chromatography, Affinity; D003429:Cross Reactions / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006168:Guinea Pigs; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000276:immunology; D009865:Oocytes / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D021122:Protein Subunits / Q000276:immunology; D018079:Receptors, GABA / Q000276:immunology*; D012160:Retina / Q000276:immunology*; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis
chemical_list: D000906:Antibodies; C087200:GABA-C receptor; D007074:Immunoglobulin G; D010446:Peptide Fragments; D021122:Protein Subunits; D018079:Receptors, GABA