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Antibodies Recognising the GABA A /Benzodiazepine Receptor Including Its Regulatory Sites
Author(s) -
Stephenson F. Anne,
Casalotti Stefano O.,
Mamalaki Cleanthi,
Barnard Eric A.
Publication year - 1986
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1986.tb13050.x
Subject(s) - muscimol , flunitrazepam , chaps , polyclonal antibodies , gabaa receptor , chemistry , receptor , immunoprecipitation , antibody , benzodiazepine , western blot , biochemistry , microbiology and biotechnology , biology , membrane , immunology , gene
Polyclonal antibodies have been raised against the GABA A /benzodiazepine receptor purified to homogeneity from bovine cerebral cortex in deoxycholate and Triton X‐100 media. Radioimmunoassay was applied to measure specific antibody production using the 125 I‐la‐belled ‐γ‐aminobutyric acid (GABA)/benzodiazepine receptor as antigen. The antibodies specifically immuno‐precipitated the binding sites for [ 3 H]muscimol and for [ 3 H]flunitrazepam from purified preparations. In addition, when a 3‐[(3‐cholamidopropyl)dimethylammonio] 1‐propanesulphonate (CHAPS) extract of bovine brain membranes was treated with the antibodies, those sites as well as the [ 3 H]propyl‐ß‐carboline‐3‐carboxylate binding, the [ 35 S]t‐butylbicyclophosphorothionate binding (TBPS), the barbiturate‐enhanced [ 3 H]fluni‐trazepam binding, and the GABA‐enhanced [ 3 H]fluni‐trazepam binding were all removed together into the immunoprecipitate. Western blot experiments showed that these antibodies recognise the α‐subunit of the purified GABA/benzodiazepine receptor. These results further support the existence in the brain of a single protein, the GABA A receptor, containing a set of regulatory binding sites for benzodiazepines and chloride channel modulators.