Polyacrylamide gel technique for the histochemical demonstration of soluble enzymes.
Soluble enzymes were immobilized and visualized by polyacrylamide gel slabs, impregnated with the incubation medium including auxiliary enzymes. The method has several advantages over existing techniques which make use of gel films or a semipermeable membrane. The diffusion of tissue compounds is effectively limited, while auxiliary enzymes may be operative. Moreover the viscosity of the medium is temperature-independent so that the incubation temperature can be varied. To demonstrate the suitability of the method glycerol-3-phosphate dehydrogenase, lactate dehydrogenase, glucose-6-phosphate dehydrogenase, hexokinase, phosphoglucomutase and aldolase were visulaized in human or rat skeletal muscle. Cytosolic and mitochondrial glycerol-3-phosphate dehydrogenase were both visualized in the absence of added NAD+ and menadione. For the visualization of ATP producint enzymes, like creatine kinase and pyruvate kinase, the method is not suitable.
D000818:Animals; D003600:Cytosol / Q000201:enzymology; D004798:Enzymes / Q000032:analysis*; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis; D005782:Gels; D005954:Glucosephosphate Dehydrogenase / Q000032:analysis; D005993:Glycerolphosphate Dehydrogenase / Q000032:analysis; D006593:Hexokinase / Q000032:analysis; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D009132:Muscles / Q000201:enzymology; D010733:Phosphoglucomutase / Q000032:analysis; D051381:Rats
D004798:Enzymes; D005782:Gels; D005993:Glycerolphosphate Dehydrogenase; D007770:L-Lactate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D006593:Hexokinase; D005634:Fructose-Bisphosphate Aldolase; D010733:Phosphoglucomutase