Summary
The detailed characterization of complex protein mixtures as in samples from biological sources cannot be sufficiently performed by separation of polypeptides according to their molecular weight as is done by conventional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (1DE). For analysis of such samples, 2-dimensional gel electrophoresis (2DE) is the preferable methodological approach because it combines separation of polypeptides according to isoelectric properties and molecular weight as well. The resulting pattern of protein spots does not only provide information on composition of samples because of the complexity of a mixture of polypeptides. It delivers also a picture on the microheterogenity of polypeptides caused by post-translational modifications. These might be of natural or artificial type and occur during biosynthetic processing of a polypeptide or within industrial scale production. The presented method describes an experimental approach to investigate the influence of glycosylation in general and sialylation exclusively on spot pattern of proteins separated by 2DE.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Löster, K., Kannicht, C. (2008). 2-Dimensional Electrophoresis: Detection of Glycosylation and Influence on Spot Pattern . In: Kannicht, C. (eds) Post-translational Modifications of Proteins. Methods in Molecular Biology™, vol 446. Humana Press. https://doi.org/10.1007/978-1-60327-084-7_14
Download citation
DOI: https://doi.org/10.1007/978-1-60327-084-7_14
Publisher Name: Humana Press
Print ISBN: 978-1-58829-719-8
Online ISBN: 978-1-60327-084-7
eBook Packages: Springer Protocols