Immobilization of Monosaccharides Fucose

Shake 15 g of the washed wet Sepharose together with 10 ml of Soln. A, 0.5 g fucose, and 75 ml pure water at 40 °C overnight. Add 0.6 g of ethanolamine and continue shaking at 40 °C for further 2 h. Remove liquid and wash with 500 ml pure water. 

Matsumoto I, Mizuno Y, Seno N (1979) J Biochem 85 1091 Sundberg L, Porath J (1974) J Chromatogr 90 87 

Carbohydrates as well as proteins are effectively coupled to divinyl-sulfone-activated agaroses. This variant is often more favorable then the activation by epoxides. As further advantage, the resulting vinyl group serves as a spacer. 

5 M Na2C03, adjusted to pH 10 with NaOH C 0.1 M ethanolamine, adjusted to pH 8 with HCl 

Wash 25 ml settled gel (e.g., Sepharose CL-4B) on a Buchner funnel with 100 ml pure water, followed by 50 ml Soln. A. Transfer the gel into a lockable flask and add 25 ml of Soln. A. Resuspend the gel and add 2.5 ml of divinyisuifone. Close the flask and agitate at RT for 1 - 2 h. Next wash the gel on a Buchner or sintered-glass funnel with 200-250 ml of Soln. B. Suck off the excess liquid. 

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