Quantitative Immunoprecipitation Studies with Anti-γ-Aminobutyric AcidA Receptor γ2 1–15 Cys Antibodies |
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Authors: | Michael J Duggan Simon Pollard F Anne Stephenson |
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Institution: | Department of Pharmaceutical Chemistry, School of Pharmacy, University of London, England. |
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Abstract: | Antibodies raised against the synthetic peptide NH2-QKSDDDYEDYASNKTC-COOH (gamma 2 1-15 Cys), which corresponds to the N-terminal amino acid sequence with a C-terminal cysteine of the human gamma 2 subunit of the gamma-aminobutyric acidA (GABAA) receptor, were used to study the quantitative immunoprecipitation of agonist benzodiazepine binding sites from bovine brain. Anti-gamma 2 1-15 Cys antibodies were found to immunoprecipitate specifically in parallel 3H]flunitrazepam- and 3H]muscimol-reversible binding sites in a dose-dependent manner. The maximum percentages of 3H]flunitrazepam binding sites immunoprecipitated from detergent extracts of bovine cerebral cortex, cerebellum, and hippocampus were 68, 77, and 83%, respectively. Immunoprecipitation studies with anti-alpha 1 324-341 antibodies carried out in parallel with anti-gamma 2 1-15 Cys antibodies provided evidence for the promiscuity of the gamma 2 subunit within native GABAA receptors. These results substantiate the association of the gamma 2 polypeptide with native GABAA receptors. |
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Keywords: | γ-Aminobutyric acid Benzodiazepine Receptor Antibody |
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