By Arthur Weissbach and Herbert Weissbach (Eds.)
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74 T. J. Waeghe, A. G. Darvill, M. McNeil, and P. Albersheim, Carbohydr. Res. 123, 281 (1983). 73 28 I. CELL WALL AND MEMBRANE is directed into each test tube. Needle valves individually regulate the amount of air introduced into each test tube. Such an apparatus is illustrated in Fig. 3 ml of isopropyl alcohol is added and evaporated. The evaporation of isopropyl alcohol results in more complete removal of the TFA. The glycoses are then reduced to the corresponding alditols by the addition of 250 μΐ of a solution of 1 M ammonia containing 10 mg/ml of sodium borohydride.
24 I. CELL W A L L AND MEMBRANE change column at much higher buffer concentrations (see Fig. 7). Isolating sycamore pectic polymers is accomplished more readily from cell walls than from SEPS. A polysaccharide similar to cell wall RG-I may be present in very small amounts in SEPS, but has not yet been isolated. An RG-II-like polysaccharide can, however, be isolated from peak C of the anion-exchange column. 2). A polysaccharide containing all the unusual sugars characteristic of RG-II (see Table II) is eluted in the partially included volume of the P-10 column.
Desorption of deesterified pectic polymers from the anion-exchange resin demands high concentrations of salt, and so yields of these molecules from anion-exchange columns tend to be very low. To avoid precipitation of the desorbed pectic polymers, the resin is eluted with a concentration gradient of imidazolium chloride. Pectic polymers are quite soluble in high concentrations of this salt. 0). 0). The EPGtreated SEPS (in the same buffer) is loaded and the column is then washed with additional buffer (1 column volume).
Methods for Plant Molecular Biology by Arthur Weissbach and Herbert Weissbach (Eds.)