Protocol

Part of the series Methods in Molecular Biology pp 1-10

Date:

Methods for Analyzing Sphingosine-1-Phosphate Signaling in Human and Mouse Primary Mast Cells

  • Alena P. ChumanevichAffiliated withDepartment of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine
  • , Piper A. WedmanAffiliated withDepartment of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine
  • , Carole A. OskeritzianAffiliated withDepartment of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine Email author 

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Mast cells produce a potently bioactive sphingolipid metabolite sphingosine-1-phosphate (S1P) constitutively and upon activation. The ligation of S1P to its type 2 receptor on mast cells triggers a novel downstream signaling pathway that we discovered links activation of transcription factor signal transducer and activator of transcription 3 to mast cell-derived chemokine release in both humans and mice. In this chapter, we describe the methods used to study S1P signaling in human and mouse primary mast cells.

Keywords:

Primary mast cells Sphingosine-1-phosphate Signaling Protein phosphorylation Signal transducer and activator of transcription 3 Activation Chemokines Inflammation Western blot Quantitative PCR