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 sensible variety of the cellphone, the difficult research of the glycome has accelerated past the study of carbohydrate specialists and into the broader scope of the lifestyles sciences. to assist all scientists now delving into this important topic region, Glycomics: equipment and Protocols collects a compendium of precise laboratory protocols reflecting the expanding availability of pattern instruction, chromatographic, electrophoretic, mass spectrometric, and bioinformatic instruments particularly designed for the research of glycosylation. major researchers within the box deal with matters equivalent to glycoprotein and proteoglycan research, glycosylation constitution choice, in addition to a number of ways to enquire the interplay among glycans and a number of carbohydrate-recognizing proteins that allows you to reduction exploration into the sensible importance of the oligosaccharides. Written within the hugely winning Methods in Molecular Biology™ sequence layout, the chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, simply reproducible protocols, and notes on troubleshooting and heading off recognized pitfalls.
Authoritative and state-of-the-art, Glycomics: equipment and Protocols serves as a worthy consultant for experimenters dealing with the demanding situations of glycan research in wish of delivering additional insights into the biology of cell-cell communique and interaction.
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Extra info for Glycomics: Methods and Protocols
Am. Chem. Soc. 117, 6783–6784. 25. Little, D. , Aaserud, D. , Valaskovic, G. A. and McLafferty, F. W. (1996) Sequence information from 42–108-mer DNAs (complete for a 50-mer) by tandem mass spectrometry J. Am. 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.
This solution has to be prepared immediately before use. 5. This solution can be stored at −20°C for several months. 6. This protocol has been optimized for 1 mg of glycoprotein, although amounts of up to 3 mg can be processed. If a larger amount of glycoprotein is to be digested, adjust the volume of reagents accordingly. 7. Use a plastic pipette for the NaOH solution and a glass syringe for the MeOH. 8. During this base preparation, a significant quantity of sodium carbonate precipitates on the wall of the tube; this must be removed as thoroughly as possible with a Pasteur pipette.
A typical pulse sequence employed for IRMPD in an FTICR mass spectrometer. 421. On the basis of the exact mass, monosaccharide compositions for each component were obtained. The sequence of each component was determined by IRMPD as shown below. Figure 3 shows a typical pulse sequence for MALDI-IRMPD analysis. The Nd:YAG laser was fired five times at 1,000-ms intervals to get enough ions in the ICR cell. A desired ion is readily selected in the analyzer with the use of an arbitrary waveform generator and a frequency synthesizer.