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

  • Barbara VisentinEmail author
  • Gary Reynolds
  • Roger Sabbadini
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 874)

Abstract

Sphingosine-1-phosphate (S1P) and the enzyme primarily responsible for its production, sphingosine kinase-1 (SphK-1), are thought to be 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 powerful tools not only to understand the 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 two staining procedures for identification of S1P and SphK-1 in human frozen tissue samples and the challenges encountered in the process of localization in tissue samples of lipid molecules, such as S1P.

Key words

Bioactive lipids S1P Sphk-1 Anti-S1P antibody LT1002 Hepatocellular carcinoma Immunohistochemistry Tumor tissue 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Barbara Visentin
    • 1
    Email author
  • Gary Reynolds
    • 2
  • Roger Sabbadini
    • 1
  1. 1.Lpath IncorporatedSan DiegoUSA
  2. 2.Centre for Liver Research, NIHR Biomedical Research UnitUniversity of Birmingham & Queen Elizabeth Hospital Birmingham NHS TrustBirminghamUK

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