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The measurement of glutamate decarboxylase activity in brain tissues by a simple microradiometric method

โœ Scribed by Joseph R. Moskal; Subhash Basu

Publisher
Elsevier Science
Year
1975
Tongue
English
Weight
507 KB
Volume
65
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

A simple method was developed for the assay of L-glutamate decarboxylase (GAD) in brain homogenate and subcel~ul~ fractions. In a dual tube assay system, the incubation mixture (0.1 ml) was placed in an inner disposable culture tube (6 x 50 mm) which was then placed in an outer reusable culture tube (10 x 75 mm) and sealed with a serum cap. The procedure utilizes absorption of '"CO, (released from [ I-"C]t-glutamate) on a Hyamine hydroxide spotted Whatman 3MM chromatographic paper strip (5 x 40 mm) which is then counted with a MiniVial containing 5 ml of toluene scintillation solution. Only very small tissue samples (0.05-0.4 mg of protein) and minimum manipulations are necessary. The assay method reported here could be used for the measurement of any decarboxyiase-catalyzed reaction, provided that suitable carboxyl labeled substrates are employed.

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