Abstract
Antibodies against gamma-aminobutyric acid (GABA)-glutaraldehyde-lysine were obtained by using a procedure based upon (i) a high yield of coupling of GABA to protein carriers, (ii) the reduction of the resulting immunoreactive double bonds, and (iii) a protocol of alternative immunizations using different immunogens having in common only the GABA-glutaraldehyde-lysine segment. This strategy led to the use of the resulting GABA antiserum without further purification. Specificity controls have been carried out with a radiolabeled ligand, [3H]GABA-glutaraldehyde- prolylphenylalanyl -lysine , which mimicked the structure of the immunogen and the fixed hapten in the tissue. Displacement curves showed that the nearest coupled analogs, beta-alanine and glycine, cross-react poorly with GABA, requiring 175-fold or 795-fold higher concentrations, respectively. Immunocytochemical results indicated that the localization obtained with this GABA antiserum largely corresponds with that reported after glutamate decarboxylase immunocytochemistry. The approach may have general applicability to other small molecules such as amino acids.
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