Abstract
The GABAC 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 GABAC 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, GABAC Ab N-14 shows reactivity with the ρ1 subunit of preparations obtained from ρ1 GABAC-expressing neuroblastoma cells, Xenopus oocytes, and mammalian retina and brain. Flow cytometry reveals a rightward shift in mean fluorescence intensity of GABAC-expressing neuroblastoma cells probed with GABAC Ab N-14. Immunostaining of neuroblastoma cells and oocytes with GABAC Ab N-14 yields fluorescence only with GABAC-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. GABAC Ab N-14 appears well suited for investigative studies of GABAC ρ1 subunit in retina and other neural tissues.
Original language | English (US) |
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Pages (from-to) | 59-64 |
Number of pages | 6 |
Journal | Experimental Eye Research |
Volume | 93 |
Issue number | 1 |
DOIs | |
State | Published - Jul 2011 |
Externally published | Yes |
Bibliographical note
Funding Information:We thank Dr. David S. Weiss for the gift of SHp5-ρ1 neuroblastoma cells; Drs. Nasser M. Qtaishat, Deborah M. Little and Brian K. Kay for helpful discussions; and Mr. Ambarish Pawar, Ms. Hongyu Ying and Dr. Raja Fayad for technical assistance. Supported by NIH grants EY016094, EY001792 and HL24530 ; by grants from the Daniel F. and Ada L. Rice Foundation (Skokie, IL) ; by the Macular Degeneration Research Program of the American Health Assistance Foundation (Clarksburg, MD) ; by Hope for Vision (Washington, DC) ; and by Research to Prevent Blindness (New York, NY) .
Keywords
- Antibody
- GABA
- GABA receptor
- Retina
- ρ1 Subunit