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Galectins pp 63-74 | Cite as

Evaluation of Galectin Binding by Frontal Affinity Chromatography (FAC)

  • Jun Iwaki
  • Jun HirabayashiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1207)

Abstract

Frontal affinity chromatography (FAC) is a simple and versatile procedure enabling quantitative determination of diverse biological interactions in terms of dissociation constants (K d), even though these interactions are relatively weak. The method is best applied to glycans and their binding proteins, with the analytical system operating on the basis of highly reproducible isocratic elution by liquid chromatography. Its application to galectins has been successfully developed to characterize their binding specificities in detail. As a result, their minimal requirements for recognition of disaccharides, i.e., β-galactosides, as well as characteristic features of individual galectins, have been elucidated. In this chapter, we describe standard procedures to determine the K d’s for interactions between a series of standard glycans and various galectins.

Key words

Oligosaccharide specificity Carbohydrate-recognition domain Comparative glycomics Dissociation constant Frontal affinity chromatography Pyridylamination 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Lectin Application and Analysis Team, Research Center for Medical GlycoscienceNational Institute of Advanced Industrial Science and TechnologyTsukubaJapan
  2. 2.Glycan Lectin Engineering Team, Research Center for Stem Cell EngineeringNational Institute of Advanced Industrial Science and TechnologyTsukubaJapan

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