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Examination of Whole-Cell Galectin Binding by Solid Phase and Flow Cytometric Analysis

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Galectins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2442))

Abstract

We have utilized simple flow cytometric and fluorescence-based solid phase assays to study the interaction of glycan binding proteins (GBP) to cell surface glycoconjugates. These methods utilize commonly employed flow cytometry techniques and commercially available streptavidin-coated microplates to immobilize various biotinylated ligands, such as glycopeptides, oligosaccharides, and whole cells. Using this approach, fluorescently labeled GBPs, in particular, members of the galectin family, can be interrogated for potential interactions with cell surface carbohydrates, including elucidation of the potential impact of alterations in glycosylation on carbohydrate recognition. Using these approaches, we present examples of flow cytometric and fluorescence-based solid phase assays to study galectin–carbohydrate interactions.

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Authors and Affiliations

  1. Orion Dagnostica, Espoo, Finland

    Anne Leppänen

  2. Joint Program in Transfusion Medicine, Department of Pathology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Connie M. Arthur & Sean R. Stowell

  3. Harvard Glycomics Center, Harvard Medical School, Boston, MA, USA

    Connie M. Arthur, Sean R. Stowell & Richard D. Cummings

Authors
  1. Anne Leppänen
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  2. Connie M. Arthur
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  3. Sean R. Stowell
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  4. Richard D. Cummings
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Corresponding author

Correspondence to Richard D. Cummings .

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Editors and Affiliations

  1. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Sean R. Stowell

  2. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Connie M. Arthur

  3. Department of Surgery Beth Israel Deaconess Medical Center National Center for Functional Glycomics, Harvard Medical School, Boston, MA, USA

    Richard D. Cummings

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Leppänen, A., Arthur, C.M., Stowell, S.R., Cummings, R.D. (2022). Examination of Whole-Cell Galectin Binding by Solid Phase and Flow Cytometric Analysis. In: Stowell, S.R., Arthur, C.M., Cummings, R.D. (eds) Galectins. Methods in Molecular Biology, vol 2442. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2055-7_11

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  • DOI: https://doi.org/10.1007/978-1-0716-2055-7_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2054-0

  • Online ISBN: 978-1-0716-2055-7

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