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Extra info for Carbohydrate Metabolism, Part B
After the addition of the epimerase, a further increase in absorbance occurs and the incubation is continued until 10-15 rain after the point where the absorbance remains constant (Fig. 1). , Cambridge, Massachusetts 02142) placed either in a warm room at 37 °, or in a blast of warm air from a hair-drier. Ultrafiltration should be complete after 5 rain. The ultrafiltrate is returned to the cell and passed through a second time to remove any traces of enzyme. 4, and the solution is stored frozen at --20 °.
The polarograms of the oxidized substrates can be classified in three types. The first type belongs to the 3-uloses, produced in high yield, from lactulose (85%), lactose (80%), lactobionic acid (75%), methyl-fl-D-thiogalactose (72%), and melibiose (65%). 50 V, and the plateau of the diffusion current is not steep (Fig. 2A). The second type is shown by the putative 3-uloses, produced in much lower yield, from glucosamine (31%), methyl-a-D-glucose (22%), methyl-fl-D-glucose (19%), maltose (17%), trehalose (17%), cellobiose (16%), glucose (14%), maltobionic acid (14%), sucrose (12%), cellobionic acid (12%), galactose (12%), leucrose (6%), glucose 1-phosphate (4%), mannose  ASSAY FOR REDUCING SUGARS 27 (4%), and p-arbutine (3%).
3) is most likely valid for any 3-keto disaccharide and Eq. (4) for any 3-keto methylhexoside. The constants for any 3-ulose in Eqs. 39 for 3-ketotrihexoses. Equation (3) is valid within the range of 1 mM to 3 ~M 3-ketolactose. The smallest measurable concentration is 1 ug/ml. Below concentrations of 10 ~M, Eln shifts from - 1 . 5 0 to - 1 . 5 6 V. Interfering Effects. Proteins, even at low concentrations after removal with chloroform, change E~ by - 0 . 0 3 V and decrease i~ for at least 15%.