Chapter title |
Immunohistochemical Detection of Sphingosine-1-Phosphate and Sphingosine Kinase-1 in Human Tissue Samples and Cell Lines
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Chapter number | 44 |
Book title |
Methods in Molecular Biology
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Published in |
Methods in molecular biology, May 2017
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DOI | 10.1007/7651_2017_44 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7412-2, 978-1-4939-7413-9
|
Authors |
Reynolds, Gary M., Visentin, Barbara, Sabbadini, Roger, Gary M. Reynolds, Barbara Visentin, Roger Sabbadini |
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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