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Immunohistochemical Detection of Sphingosine-1-Phosphate and Sphingosine Kinase-1 in Human Tissue Samples and Cell Lines

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Sphingosine-1-Phosphate

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1697))

Abstract

Sphingosine-1-phosphate (S1P) and the enzyme primarily responsible for its production, sphingosine kinase-1 (SphK-1), are dysregulated in multiple human diseases including cancer, multiple sclerosis (MS), diabetes, neurological diseases, fibrosis, and certain pathologies associated with impaired angiogenesis such as age-related macular degeneration (AMD). Antibody-based techniques to identify and localize S1P and SphK-1 within cells and tissue specimens represent a powerful tool, not only to understand biological role of these molecules but also to validate these unique in-class targets in multiple state diseases. Consequently, the potential applications of these molecules for therapy and diagnostic purposes are currently under investigation. Here, we describe a new improved technique, Agitated Low Temperature Epitope Retrieval (ALTER) for staining procedures, to identify expression of S1P and SphK-1 in human frozen tissue samples. The challenges encountered in the process of localization in tissue samples of lipid molecules such as S1P are discussed.

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Author information

Authors and Affiliations

  1. Centre for Liver Research and NIHR Biomedical Research Unit, University of Birmingham and Queen Elizabeth Hospital Birmingham, Birmingham, B15 2TT, UK

    Gary M. Reynolds

  2. La Jolla Biologics Inc, San Diego, CA, USA

    Barbara Visentin

  3. Stanford University School of Medicine, Stanford, CA, USA

    Roger Sabbadini

Authors
  1. Gary M. Reynolds
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  2. Barbara Visentin
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  3. Roger Sabbadini
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Corresponding author

Correspondence to Gary M. Reynolds .

Editor information

Editors and Affiliations

  1. Centre for Eye Research Australia, The University of Melbourne, East Melbourne, Victoria, Australia

    Alice Pébay

  2. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

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Reynolds, G.M., Visentin, B., Sabbadini, R. (2017). Immunohistochemical Detection of Sphingosine-1-Phosphate and Sphingosine Kinase-1 in Human Tissue Samples and Cell Lines. In: Pébay, A., Turksen, K. (eds) Sphingosine-1-Phosphate. Methods in Molecular Biology, vol 1697. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_44

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  • DOI: https://doi.org/10.1007/7651_2017_44

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7412-2

  • Online ISBN: 978-1-4939-7413-9

  • eBook Packages: Springer Protocols

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