Histo-electrophoretic analysis of the microheterogeneity and region-specificity of isozymes.
An optimal and practical method for high resolution separation and visualization of soluble and membrane-binding isoenzymes is described. By use of this method, non-specific esterase, lactate dehydrogenase and acid phosphatase from different tissues of mice can be separated into more than 50, 30 and 20 components, respectively. Cholinesterase, glucose-6-phosphate dehydrogenase, acid phosphatase, succinate dehydrogenase, lactate dehydrogenase and non-specific esterase isolated from the lumbar spinal cord of chick embryos at different ages can be separated into about 15, 12, 9, 6, 14 and 20 components, respectively. In addition, an attempt has been made to evaluate quantitatively by densitometry differences in enzyme activity among different regions of the same tissue (e.g. spinal cord). The findings revealed that there are differences in isoenzyme components between the dorsal and ventral regions of the spinal cord. Collectively, the combination of direct tissue isoelectric focusing and fast enzyme visualization reveals a greater number of isoenzyme components than can be demonstrated by other means. This method is an extremely useful procedure for understanding and analyzing the nature and topographic localization of isoenzyme components.
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Department of Neurobiology and Anatomy, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27103.
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D000135:Acid Phosphatase / Q000302:isolation & purification; D000818:Animals; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification; D002642:Chick Embryo; D002802:Cholinesterases / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification; D007202:Indicators and Reagents; D007525:Isoelectric Focusing / Q000379:methods; D007527:Isoenzymes / Q000302:isolation & purification*; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009928:Organ Specificity; D013116:Spinal Cord / Q000201:enzymology; D013385:Succinate Dehydrogenase / Q000302:isolation & purification
D007202:Indicators and Reagents; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D013385:Succinate Dehydrogenase; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D002802:Cholinesterases; D000135:Acid Phosphatase