Measurement of erythrocyte glucose-6-phosphate dehydrogenase activity using a centrifugal analyzer.
نویسنده
چکیده
We describe two improved methods for determination of erythrocyte glucose-6-phosphate dehydrogenase activity. One method is an “enzyme-linked” procedure in which an excess of 6-phosphogluconate dehydrogenase is used to produce two moles of NADPHfor each mole of glucose-6-phosphate oxidized. In the other method 2,3-diphosphoglycerate is used to inhibit the variable contribution of endogenous 6-phosphogluconate dehydrogenase to glucose-6-phosphate dehydrogenase activity in erythrocyte lysates. These assays require 100 tI of blood and are performed on a centrifugal analyzer in a final reaction volume of 410 0 at 37 #{176}C. NADPH formation is monitored at 340 nm. Hemoglobin is measured as oxyhemoglobin in the same reaction mixture used to determine enzyme activity by changing the wavelength to 550 nm. Results are expressed as international units of glucose-6-phosphate dehydrogenase activity per gram of hemoglobin. The coefficient of correlation between “enzyme-linked” assay and “standard” assay was .992, the slope of the regression line was 1.07, and the intercept was at -0.76. When results of the “enzyme-linked” assay were compared to those of the “nonlinked” assay with added 2,3-diphosphoglycerate, the slope of the regression line was .994, the intercept 0.109, and the correlation coefficient .994.
منابع مشابه
Maleimide as an inhibitor in measurement of erythrocyte glucose-6-phosphate dehydrogenase activity.
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عنوان ژورنال:
- Medical laboratory sciences
دوره 41 2 شماره
صفحات -
تاریخ انتشار 1975