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A Laboratory Guide to Glycoconjugate Analysis by Uwe D. Neue, Charles H. Phoebe (auth.), Dr. Peter Jackson,

By Uwe D. Neue, Charles H. Phoebe (auth.), Dr. Peter Jackson, Prof. John T. Gallagher (eds.)

18. 2 precept of FACE/Gel Retardation Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 349 18. three Labelling of Oligosaccharides with ANTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 350 18. four Screening of Carbohydrate Ligands for Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . 352 18. five dimension of Binding consistent for the interplay among Protein and ANTS-Labelled Carbohydrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 355 18. 6 dimension of Binding consistent for the interplay among Protein and local Carbohydrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360 ~ the applying of Capillary Affinity Electrophoresis to the research _ of Carbohydrate-Protein Interactions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 19. 1 creation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 19. 2 precept of CAE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363 19. three choice of organization Constants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364 19. four Technical approaches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 normal concerns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 19. five obstacles of the approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 370 19. 6 program of CAE to the research of Carbohydrate-Protein Interactions . . . . . . 371 19. 7 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 375 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 377 20. 1 creation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 379 Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 380 20. 2 Technical strategies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 381 20. three pattern Detection and pattern restoration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Autoradiography and marking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 pattern detection via blotting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Semipreparative ACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 390 20. four research of information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 391 Measuring pattern mobilities - calculating a retardation coefficient . . . . . . . . . . . . 391 Graphical research of knowledge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 392 studying ACE styles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393 opposite ACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395 20. five precis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 397 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 topic Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399 XII record of members Nebojsa Avdalovic John T. Gallagher Dionex company melanoma examine crusade division of clinical Oncology 445 Lakeside force college of Manchester Sunnyvale, CA 94086 Christie CRC learn Centre Klaus Biemann Wilmslow highway division of Chemistry Manchester M20 4BX Massachusetts Institute of know-how united kingdom Cambridge, MA 02139-4307 united states Geoffrey R.

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The first publication also includes an overview of the theory and operating conditions 19 of immobilized enzyme reactors in general and is a good starting point for researchers who would like to investigate similar techniques. References I Journal oJ Chromatography A 720 (1996) 2 Edge CJ, Rademacher TW, Wormald MR, Parekh RB, Butters TD, Wing DR and Dwek RA (1992) Proc. Natl. Acad. Sci. USA 89: 6338 3 Green ED and Baenziger JU (1986) Anal. Biochem. 158: 42 4 5 Damm JBL, Kamerling JP, van Dedem 8 J.

As with most separation techniques based on adsorption, one obtains a separation of the anomers, which makes the chromatogram quite crowded. 75 mllmin. At room temperature and neutral pH two peaks are obtained for each oligomer due to the separation of the anomers. Anomer separation can be avoided by using elevated temperature, derivatization or best with high-pH mobile phases. 0 ml Imin. Peak designation according to the degree of polymerization. ative fractions. The interconversion of the anomers can be speeded up by increasing the temperature, but this happens at the expense of reduced retention times.

Soc. Trans. 20, 264. 2 Varki A (1993) Glycobiology 3: 97. \0 Laemmli UK (1970) Nature 277: 680. D. Hannes and D. 4 of the sugar chains of glycoproteins. Eur. Rickwood (eds) An Introduction to Polya- 1. Biochem. 209: 483. cylamide Gel Electrophoresis: A Practical Dwek RA, Edge Cl, Harvey 01, Wormald Approach, IRL, Oxford. MR and Parekh RB (1993) Glycoform analysis of glycoproteins. Ann. Rev. Biochem. 62: 6. 5 Kobata A (1992) Structures and functions of the sugar chains of glycoproteins. Eur. J. Biochem.

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