Electrophoresis used in blood cell enzyme polymorphism study
1997
Karus, V. | Karus, A. | Lepiku, T. (Estonian Agricultural Univ., Tartu (Estonia))
At present a lot of the genetic markers, such as blood groups, protein polymorphic forms e.c. are used to solve the questions concerning animal breeding. Enzymes are very prospective study objects for acquiring of knowledge about processes in an animal organism. Recently we studied SDH and ALD isoenzyme spectra in blood serum, but there are many nongenetical influences on the isoenzyme spectra in serum. It is more prospective to investigate isoenzyme spectra in blood cells. In this study we investigated the methods for determination of isoenzyme spectra SDH, ASAT, SOD, IDH and GDH in serum and erythrocytes. We used native 7,5 per cent, 10,0 per cent and 12,5 percent PAGE electrophoresis and isoelectrical focusing in standard horizontal Ampholine pH 3.5-9.5 gels and vertical 5 per cent PAGE. The best results were obtained by isoelectrical focusing. The standard horizontal Ampholine pH 3.5-9.5 gel, was preferred because a linear gradient of pH in the gel, although, resolution of isoenzymes was sufficient in native 7.5 per cent and 10.0 per cent PAGE. Histochemical methods for enzyme determination gave satisfactory results. Dark background was obtained only by ASAT staining, and it makes impossible to detect the other enzymes in the same gel. Staining after IEF needed 1.5-times more concentrated staining puffer and at longer staining time
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