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 size of Binding consistent for the interplay among Protein and ANTS-Labelled Carbohydrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 355 18. 6 size of Binding consistent for the interplay among Protein and local Carbohydrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360 ~ the appliance of Capillary Affinity Electrophoresis to the research _ of Carbohydrate-Protein Interactions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 19. 1 creation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361 19. 2 precept of CAE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363 19. three selection of organization Constants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364 19. four Technical approaches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 normal issues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 19. five boundaries of the procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 370 19. 6 program of CAE to the research of Carbohydrate-Protein Interactions . . . . . . 371 19. 7 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 375 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 377 20. 1 advent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 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 analyzing ACE styles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393 opposite ACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395 20. five precis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 397 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 topic Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399 XII checklist of members Nebojsa Avdalovic John T. Gallagher Dionex company melanoma learn 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 expertise united kingdom Cambridge, MA 02139-4307 united states Geoffrey R.

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The solubility of the analytes in the mobile phase (100% water) is excellent, and no additional cleanup steps are needed after the chromatographic step. Also, large-diameter columns are readily available if the preparation needs to be scaled up to larger amounts. In order to solve the problems associated with anomer separation, the sample can be reduced prior to the chromatography. However, this may limit the usefulness of the isolated oligosaccharide for further experiments. Many different types of RP packings are commercially available.

If a signal is detected, this usually indicates that the sample contains carbohydrate; however, false positive results owing to the following causes need to be considered: • An artefact has been generated by an interfering substance incorporating biotin. • Other nonspecific binding of signal-generating compounds has occurred. • Reaction of the biotin hydrazide with noncarbohydrate aldehyde groups has occurred. However, solution labelling is extremely sensitive and may give positive responses for proteins which are not conventionally glycosylated, such as glycanated proteins.

Biochem. 30: DF, Turler Hand Balant LP (1994) 148. Anal. Biochem. 221: 66. Bayer EA, Ben-Hur Hand WiIchek M 15 Brummell DA, Sharma VP, Anand NN, (1990) Methods Enzymol. 184: 415. Bilous D, Dubuc G, Michniewicz 1, Durrant I, Benge LC, Sturrock C, Devenish Mackenzie CR, Sadowska 1, Sigurkjold AT, Howe R, Roe S, Moore M, Scozzafava BW, Sinnat B, Young M, Bundle DR and G, Proudfoot LM and Richardson TC, Narang SA(l993) Biochemistry 32: 1180. (1990) Biotechniques 8: 564. 16 Whiteheart SW, Passaniti A, Reichner JS, Holt GD, Haltiwanger RS, and Hart OW (1989) Methods Enzymol.

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