Analysis of guanase by agarose gel electrophoresis and activity staining.
A method was developed to separate guanase by agarose gel electrophoresis and to detect its activity by staining of the bands with a mixture of the enzymes xanthine oxidase, catalase, and aldehyde dehydrogenase, the coenzyme NADP+, and a substrate of guanine, ethanol, phenazine methosulfate, nitrotetrazolium blue, and KCN in Tris-(hydroxymethyl)methylamine buffer (pH 8.0). Serum samples showed bands 1 (faster moving) and 2 corresponding to the positions of albumin and alpha 2-globulin, respectively, found by serum protein staining. The same bands were detected with guanase from human liver and kidney, although band 2 from the latter samples was not as distinct as that from the liver samples.
Nishikawa
Y
Y
Fukumoto
K
K
Watanabe
F
F
D000444:Aldehyde Dehydrogenase; D000619:Aminohydrolases / Q000302:isolation & purification*; D002374:Catalase; D004587:Electrophoresis, Agar Gel; D006148:Guanine Deaminase / Q000097:blood / Q000302:isolation & purification*; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D013194:Staining and Labeling; D014969:Xanthine Oxidase
D002374:Catalase; D014969:Xanthine Oxidase; D000444:Aldehyde Dehydrogenase; D000619:Aminohydrolases; D006148:Guanine Deaminase