Determination of glutamate decarboxylase by high-performance liquid chromatography |
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| Institution: | 1. Max Planck Institute for Physics, Föhringer Ring 6, 80805 Munich, Germany;2. Departamento de Física Teórica and Instituto de Física Teórica UAM-CSIC, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain;1. Department of Radiology, Brigham and Women''s Hospital, Harvard Medical School, Boston, MA, USA;2. Center for Pain and the Brain, Boston Children''s Hospital, Harvard Medical School, Boston, MA, 02115, USA;3. Department of Anesthesia, Perioperative, and Pain Medicine, Boston Children''s Hospital, Harvard Medical School, Boston, MA, 02115, USA;4. Department of Neurobiology, Harvard Medical School, Boston, MA, USA;1. Zhejiang University, College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Hangzhou 310058, People’s Republic of China;2. Institute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Jinan 250100, People’s Republic of China |
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| Abstract: | An improved method for the determination of glutamate decarboxylase (GAD) activity is described. The enzyme was evaluated by incubation with glutamic acid (l-Glu) in the presence of pyridoxal 5 ′-phosphate (PLP): the γ-aminobutyric acid (GABA) formed was derivatized to PTC-GABA; the latter was subsequently separated and assayed by isocratic HPLC (LiChrospher RP-18 column; isocratic elution with pH 5.8 acetate buffer in acetonitrile-water) with UV absorbance detection at 254 nm. The method described is a sensitive, reproducible and specific assay useful for following variations of GAD activity in vitro; this assay was subsequently used for the evaluation of GAD activity variations after irradiation with low doses of HeNe laser radiation. |
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