Antibodies to the rat beta 3 subunit of the gamma-aminobutyric acid A receptors.
Polymerase chain reaction was used to amplify the cDNA region that codes for the large intracellular loop of the beta 3 subunit of the gamma-aminobutyric acidA/benzodiazepine receptors (GABAAR/BZDR) from rat brain. The amplified cDNA was inserted into the prokaryotic expression vector pGEX-3X and a fusion protein containing glutathione-S-transferase and beta 3 intracellular loop moieties was expressed in bacteria. The fusion protein was affinity-purified and it was used to raise a rabbit anti-beta 3 antiserum. The anti-beta 3 antiserum immunoprecipitated the gamma-aminobutyric acidA receptor from rat and bovine brain. Immunoblots of the affinity-purified GABAAR/BZDR from bovine brain revealed that the anti-beta 3 antiserum reacted with a 57 kDa peptide, whereas the monoclonal antibody 62-3G1 that recognized both beta 2 and beta 3 reacted with 55 and 57 kDa peptides. The anti-beta 3 antiserum showed specificity for the beta 3 subunit vs beta 2 and beta 1.
Fernando
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LP
Division of Molecular Biology and Biochemistry, School of Biological Sciences, University of Missouri-Kansas City 64110-2499.
Khan
Z U
ZU
De Blas
A L
AL
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000235:genetics; D002531:Cerebellum; D002540:Cerebral Cortex / Q000276:immunology*; D006624:Hippocampus; D008969:Molecular Sequence Data; D009118:Muscimol / Q000494:pharmacology; D009830:Olfactory Bulb; D011233:Precipitin Tests; D011817:Rabbits; D051381:Rats; D018079:Receptors, GABA / Q000276:immunology*; D017421:Sequence Analysis
D000906:Antibodies; D018079:Receptors, GABA; D009118:Muscimol