By Victor Ginsburg (Eds.)
The seriously acclaimed laboratory average, Methods in Enzymology, is likely one of the such a lot hugely revered courses within the box of biochemistry. for the reason that 1955, every one quantity has been eagerly awaited, often consulted, and praised by way of researchers and reviewers alike. The sequence comprises a lot fabric nonetheless appropriate this day - actually a vital ebook for researchers in all fields of lifestyles sciences
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Leitler, J. Biol. Chem. 257, 906 (1982). 14 T. J. Dodge, C. Mitchell, and D. J. Hananhan, Arch. Biochem. Biophys. 100, 119 (1963). is R. D. C u m m i n g s , A. Srderquist, and G. Carpenter, J. Biol. Chem. 260, 11944 (1985). ~6 p. F r e d m a n , N. D. Richert, J. L. Magnani, M. C. Willingham, I. Pastan, and V. Ginsburg, J. Biol. Chem. 258, 11206 (1983). 32 ANALYTICALMETHODS  Lectin Affinity C h r o m a t o g r a p h y of Glycolipid-Derived Oligosaccharides Glycopeptides and glycoprotein-derived oligosaccharides3 or free oligosaccharides ~,2 have been the subjects of most lectin affinity chromatographic techniques.
Bidlingmeyer, S. A. Cohen, and T. L. Tarvin, J. Chromatogr. 336, 93 (1984). 37 p. W. Cheng, Anal. Biochem. 167, 265 (1987). 3s T. Bhatti, R. E. Chambers, and J. R. Clamp, Biochim. Biophys. Acta 222, 339 (1970). 39 M. R. Hardy, R. R. Townsend, and Y. C. Lee, Anal. Biochem. 170, 54 (1988). 4o D. Aminoff, Biochem. J. 81, 384 (1961). [l] TRACHEOBRONCHIAL MUCIN GLYCOPROTEINS 15 Sulfate is determined by the barium chloranilate procedure of Spencer4~ after hydrolysis for 6 hr in l NHC1 at 100 ° or by the rhodizonate method of Silvestri et al.
Aliquots of the fractions were assayed for radioactivity. 34 ANALYTICAL METHODS  oligosaccharides in solution. In other assays where multivalent oligosaccharities are presented to the lectin, the specificity appears less restricted. For example, HP lectin will form a precipitin band with ganglioside GM2 in immunodiffusion assays, 2° and ~25I-labeled HP lectin binds to GM2 and asialo-GM2 on thin-layer chromatograms. '7 The immobilized WF lectin column retarded the elution of 3H-labeled oligosaccharides derived from Forssman glycolipid (Fig.