Analysis

New PDF release: A Laboratory Guide to Glycoconjugate Analysis

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 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 decision of organization Constants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364 19. four Technical tactics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 basic concerns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 19. five boundaries of the strategy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 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 methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 381 20. three pattern Detection and pattern restoration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Autoradiography and marking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 pattern detection by means of blotting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 Semipreparative ACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 390 20. four research of knowledge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 391 Measuring pattern mobilities - calculating a retardation coefficient . . . . . . . . . . . . 391 Graphical research of information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 392 reading ACE styles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393 opposite ACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395 20. five precis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 397 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398 topic Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399 XII checklist of participants Nebojsa Avdalovic John T. Gallagher Dionex company melanoma examine crusade division of clinical Oncology 445 Lakeside force collage of Manchester Sunnyvale, CA 94086 Christie CRC study 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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I, Elsevier, Amsterdam. 9 Haltiwagner RS, Kelly WG, Blomberg MA, Dong D, Roquermone E and Hart GW (1992) Biochem. 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.

Thur;ngiens;s (Oxford GlycoSystems cat. no. GPI-02) and the native enzyme from T. bruce; (Oxford GlycoSystems cat. no. GPI-03) are commercially available. 2. Separate the cleaved product by extraction into a detergent such as Triton X-114. In this technique the uncleaved material with the lipid anchor attached will extract into the detergent phase. whereas the cleaved material will extract into the aqueous phase. 3. Once extracted, detect the exposed GPI-associated epitope CRO by the use of a suitable antibody.

Biochern. 57, 285. 19 Bordier CJ (1981) BioI. Chern. 256: 1604. 20 Kelly WG and Hart GW (1989) Cell 57: 243. 21 Kearse KP and Hart GW (1991) Proc. Natl. Acad. Sci. USA 88: 1701. 22 Holt GD, Haltiwanger RS, Torres CR and Hart GW (1987) 1. Bioi. Chern. 262: 14847. 23 Alexander et al (1989) Methods Enzyrnol. 179: 505. 24 Glasgow LR and Luecke RH (1977) 1. BioI. Chern. 252: 8615.

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