Methods for studying the glucose-6-phosphate dehydrogenase activity in brain areas

被引:20
|
作者
Ninfali, P
Aluigi, G
Pompella, A
机构
[1] Univ Urbino, Ist Chim Biol G Fornaini, I-61029 Urbino, Italy
[2] Univ Siena, Ist Patol Gen, I-53100 Siena, Italy
来源
BRAIN RESEARCH PROTOCOLS | 1997年 / 1卷 / 04期
关键词
glucose-6-phosphate dehydrogenase; cytochemical assay; spectrophotometric assay; electrophoretic pattern; brain area;
D O I
10.1016/S1385-299X(97)00011-1
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This paper reports on the protocols for the spectrophotometric determination of the enzymatic activity, the electrophoretic pattern and the cytochemical assay of glucose-6-phosphate dehydrogenase (G6PD) in different areas of rat brain. For the spectrophotometric assay we used the method of Glock and McLean. Non-denaturing polyacrylamide gel electrophoresis was performed using 1.5 mm thick slab-gel, with a 7.5% acrylamide separating gel and a 4% acrylamide stacking gel. NADP (0.01 mM) was included at the cathode buffer. The cytochemical assay was performed on unfixed cryostat sections, by incubating the slides for 5 min at 37 degrees C in the staining medium. A combination of these techniques offers a means for the study of the effect of age, drugs, oxidizing or reducing agents on the G6PD activity in different brain areas. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:357 / 363
页数:7
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