By Willy Morelle, Valegh Faid, Frédéric Chirat, Jean-Claude Michalski (auth.), Niclas G. Karlsson, Nicolle H. Packer (eds.)
Due to the numerous contributions of carbohydrates to the practical range of the cellphone, the not easy examine of the glycome has elevated past the study of carbohydrate specialists and into the broader scope of the existence sciences. to assist all scientists now delving into this very important topic quarter, Glycomics: equipment and Protocols collects a compendium of unique laboratory protocols reflecting the expanding availability of pattern training, chromatographic, electrophoretic, mass spectrometric, and bioinformatic instruments particularly designed for the research of glycosylation. major researchers within the box handle matters comparable to glycoprotein and proteoglycan research, glycosylation constitution choice, in addition to quite a few techniques to enquire the interplay among glycans and quite a few carbohydrate-recognizing proteins which will relief exploration into the useful value of the oligosaccharides. Written within the hugely profitable Methods in Molecular Biology™ sequence structure, the chapters comprise introductions to their respective subject matters, lists of the mandatory fabrics and reagents, step by step, with no trouble reproducible protocols, and notes on troubleshooting and warding off recognized pitfalls.
Authoritative and state-of-the-art, Glycomics: equipment and Protocols serves as a helpful advisor for experimenters dealing with the demanding situations of glycan research in wish of delivering additional insights into the biology of cell-cell conversation and interaction.
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15. 20 M sodium carbonate to the sample. Vortex. 16. Add approximately 2 mL of methylene chloride to the sample and mix gently. 17. 4). 18. Dissolve the dried sample in 10–15 drops of methylene chloride and inject 1 μL onto the GC-MS. 6. Gas Chromatography-Mass Spectrometry (Fig. 2) Any commercially available GC-MS instrument can be used for the analysis of partially methylated alditol acetates. We use a Hewlett Packard 5890 GC interfaced to a 5970 mass selective detector. Perform the separation on a 30 m Supelco 2330 bonded phase fused silica capillary column using the following temperature program: After 2 min at the initial temperature of 80°C, ramp to 170°C at a rate of 30°/min, then ramp to 235°C at rate of 4°/min and hold at 235°C for 20 min.
Chem. Soc. 118, 9352–9359. 35 26. Little, D. P. and McLafferty, F. W. (1996) Infrared photodissociation of non-covalent adducts of electrosprayed nucleotide ions. J. Am. Soc. Mass Spectrom. 7, 209–210. 27. Xie, Y. and Lebrilla, C. B. (2003) Infrared multiphoton dissociation of alkali metal-coordinated oligosaccharides. Anal. Chem. 75, 1590–1598. 28. , Russell, S. and Lebrilla, C. B. (2005) Infrared multiphoton dissociation of O-linked mucintype oligosaccharides. Anal. Chem. 77, 208–214. 29. , Li, B.
IRMPD spectrum of an O-linked oligosaccharide with m/z 1,122 ([M + Na]+, see Fig. 2). 7 W. The primary component sequence of the O-linked glycan was determined based on the spectrum and shown in the inset. Filled circle Hex; open square GalNAc-ol; open triangle dHex; crossed square HexNAc. Infrared Multiphoton Dissociation Mass Spectrometry 31 by the loss of the other internal Hex. There is another HexNAc loss (m/z 408) from the ion m/z 611, which comes directly from m/z 814 through the loss of one HexNAc, thus indicating that the two HexNAc are connected to each other.