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

doi:10.1016/j.exer.2011.04.009

Subunit-specific polyclonal antibody targeting human ρ1 GABA_C 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 journal › Article › peer-review

2 Scopus citations

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.

Original language	English (US)
Pages (from-to)	59-64
Number of pages	6
Journal	Experimental Eye Research
Volume	93
Issue number	1
DOIs	https://doi.org/10.1016/j.exer.2011.04.009
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) .

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Antibody
GABA
GABA receptor
Retina
ρ1 Subunit

Publisher link

10.1016/j.exer.2011.04.009

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title = "Subunit-specific polyclonal antibody targeting human ρ1 GABAC receptor",

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.",

keywords = "Antibody, GABA, GABA receptor, Retina, ρ1 Subunit",

author = "Gussin, \{H{\'e}l{\`e}ne A.\} and Khasawneh, \{Fadi T.\} and An Xie and Feng Feng and Adnan Memic and Haohua Qian and \{Le Breton\}, \{Guy C.\} and Pepperberg, \{David R.\}",

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) .",

year = "2011",

month = jul,

doi = "10.1016/j.exer.2011.04.009",

language = "English (US)",

volume = "93",

pages = "59--64",

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T1 - Subunit-specific polyclonal antibody targeting human ρ1 GABAC receptor

AU - Gussin, Hélène A.

AU - Khasawneh, Fadi T.

AU - Xie, An

AU - Feng, Feng

AU - Memic, Adnan

AU - Qian, Haohua

AU - Le Breton, Guy C.

AU - Pepperberg, David R.

N1 - 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) .

PY - 2011/7

Y1 - 2011/7

N2 - 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.

AB - 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.

KW - Antibody

KW - GABA

KW - GABA receptor

KW - Retina

KW - ρ1 Subunit

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DO - 10.1016/j.exer.2011.04.009

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SN - 0014-4835

VL - 93

SP - 59

EP - 64

JO - Experimental Eye Research

JF - Experimental Eye Research

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