Purification of L-Glutamate decarboxylase from rabbit brain and preparation of a monospecific antiserum

Publisher Website
Google Scholar
Abstract
L‐Glutamate decarboxylase (GAD), the enzyme responsible for the biosynthesis of gamma‐aminobutyric acid, has been purified from rabbit brain and used for the production of a monospecific rabbit antiserum. The enzyme was purified approximately 500‐fold by a combination of ion exchange, hydrophobic, hydroxyapatite, and gel filtration column chromatography, and preparative polyacrylamide gel electrophoresis. The preparation thus obtained was injected into a rabbit, producing a polyspecific antiserum. This first antiserum was used for the preparation of an immunoprecipitate containing GAD; injections of the immunoprecipitate into a second rabbit led to the production of a second, monospecific anti‐GAD serum. The specificity of the antiserum for GAD has been demonstrated by enzyme precipitation, immunodiffusion, line immunoelectrophoresis, gel filtration chromatography of GAD/anti‐GAD immune complexes, and immunocytochemistry in the rat cerebellum.