Subunit-specific polyclonal antibody targeting human ρ1 GABAC receptor

Hélène A. Gussin, Fadi T. Khasawneh, An Xie, Feng Feng, Adnan Memic, Haohua Qian, Guy C. Le Breton, David R. Pepperberg

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

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 languageEnglish (US)
Pages (from-to)59-64
Number of pages6
JournalExperimental Eye Research
Volume93
Issue number1
DOIs
StatePublished - Jul 1 2011
Externally publishedYes

Keywords

  • Antibody
  • GABA
  • GABA receptor
  • Retina
  • ρ1 Subunit

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